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ghiajake
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Liquid Culture De-Mystified: Ghia Style 3
#19541843 - 02/08/14 09:47 PM (9 years, 11 months ago) |
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De-vilifying an extremely useful and easy tool: by GhiaJake Whether you are new or old to the debate between agar and LC (liquid culture), everyone can agree that LC has a bad rep here. Opponents of LC always state that it just isn't worth it and you would be spending your time better just using agar instead. I don't see how so many people can just blatantly dismiss such a valuable tool in mycology, other than people giving up after too many failed attempts. Anyone that has ever bought a culture syringe from a sponsor is using LC. If agar was SO much better than LC, then why don't the sponsors all sell wedges instead? There is no better way to grow, and store, mass amounts of usable culture in such a small area as with LC. Period. Yes, agar is great for germinating spores and isolating genetics. But how many dishes would it take to inoculate over 200 jars of grain. One quart jar with only 600mL of LC can do it easily. Also, when storing agar dishes in the fridge the moisture will separate from the agar, leaving pools of water lying on your culture. The LC culture is always hydrated, never get opened to the air (sterile or otherwise), and just continues to grow while being stored. I went through a patch of not wanting to mess with agar, since I got reports of contaminated wedges I had traded. Again, to anyone that received a contaminated wedge, I'm sorry. To be honest, I still haven't done any new agar work since, even since I built my flow hood. I decided instead to spend my time mastering the "dreaded" LC. I have to give credit to Claybuddy for showing me that LC is viable and should be looked into more (How else are you going to grow non-sporulating Purple Mystics, right?  ). I can't remember the link he gave me to where he learned his LC recipe, and I by no means claim it to be mine, but below I will describe how I now do my LC jars. To start off, here's a few pics of some of the LCs I've made using my method. The first is Shiitake (S75) inoculated with sterile dowel spawn. The second is King Oyster inoculated with a few colonized milo grains. The third is Pearl Oyster (or pseudo Hyp Ulm) noc'd with dowel spawn. The forth is Ps galindoi, noc'd with 5cc of GLC. The fifth is Pan cyan noc'd the same as the ATL#7.      Now, enough bragging, let's get down to business making some LC! Keep in mind, LC is meant for live culture transfers only!!! Trying to germinate spores in an LC solution will end in failure most of the time. There are many ways people attempt to do the whole LC thing. I can say I've tried most of them by now and this is the easiest method, with the best results. There are a few things you are going to need to pick up to do this right. Keep in mind that if you half-ass things you will only ever get half-ass results. Spending the extra time to do things right will be well worth it in the end. Here's what you will need to make your "no-tilt" jar lids:  Plastic jar lids Self-healing inoculation ports "Whatman-style" syringe filters (PVDF or PTFE, 0.2 micron) Rubber grommets (tattoo gun grommets work fine, but I like the thicker ones) 1/4" stainless tubing 1/2" drill bit 1/4" drill bit (3/8" if you use the thicker grommets) High-temp silicone (optional, not really needed as the tubing seals the noc port) Step 1: Step 2: Start off by drilling two holes (carefully) in each lid with your two drill bit sizes. Clean the edges of the holes with a knife, or whatever. Step 3:  Insert the noc port into the 1/2" hole, it should snap into the retention slot on the noc port. The grommet fits in the 1/4" hole. You can see the difference between the two grommets in the above picture. Step 4: Carefully insert the filter syringe through the grommet. If you use the tattoo gun grommets, you will want to watch that you don't push the grommet through the hole. Make sure the filter and grommet are properly seated to the lid. You can use your high-temp silicone for added protection at this time, but it isn't really needed when making your lids this way. Step 5:   It is important to get stainless steal tubing for your "downspout". You don't want rust in your LC! For quart sized jars cut your length to 6" from base to tip, 4" for pint jars. Cut one end square, and the other at a drastic angle. This allows for a much larger area of myc to be drawn into the tubing. I also sharpen the edges of the tips to help with cutting up the mycelium when swirling (see section on swirling) Smooth the edge on the other end, and slip it into the bottom of noc port. It will fit quite snugly, and will keep the noc port from pulling back out of the lid (see above pic) Make sure to turn the tip of your downspout into the direction of your swirl. You want the sharpened tip to cut the oncoming mycelium when it is spinning in the jar Step 6:  Now that you have your lids made, you're almost ready to start on the LC itself. If you don't have a magnetic stirrer, just toss in a couple clean stones into each of your jars. Step 7:  BTW, from here on I'll have to update later with photos. I have a batch to make in the next couple of days, so I'll update the OP once I get all that done. For now, we continue on... Step 8: Now that all your jars are ready, it's time to mix up your liquid. This is one of those widely debated subjects, which nutrient base to use in your LC. I have tried many different ways: honey and/or karo, potato water, grain soak water, etc. The best results I've found are from just extra light malt extract. It's cheap, has great results, and you barely need any for it to work. I use 0.6g of ME(got it by mistake) for every 100mL of DISTILLED water. Don't use tap water folks! Miss a beer and go buy a gallon of distilled water for Jebus' sake! Or use water collected in a house dehumidifier like I do.  What I usually do is pour all the water into a big pot and bring it to a boil. Shut off the stove, then stir in your malt extract. Make sure it all dissolves. I then pour the brew through a double-layered clean t-shirt to filter out the particulates. Do this until your liquid is clear. Now fill up your jars and get the PC ready. Double check that you did both Step 5 and Step 6 before closing your lids, since Step 7 sometimes makes you forget things. Step 9: Pressure cook your LC jars at 15-18 PSI for 30 minutes. There is no need for aluminum foil over the lids. Make sure you don't quickly release the pressure at any point or your jars will expel your LC liquid due to the pressure equalization. Just let them cool down in the PC until the next morning. Your jar temps need to be below 80F before inoculating anyways. If you correctly filtered the liquid before pressure cooking, you shouldn't have any particulates in your jars after PC'ing. If you have some, don't worry. The mycelium should eat it. You may notice a color change if you use ME instead of ELME, but again don't worry. It's fine. Step 10: Now we're onto the meat of the issue. Inoculation. The whole agar thing and I can't see eye-to-eye most of the time (fucking agar), so I do my LC a little different than most. If you have no problems with agar then just noc up your LC with it if you like. But this way you aren't "touching" the culture in any way, so you are eliminating that chance of contamination All inoculation methods should be done in a SAB, or in front of a laminar hood, and all sterile procedures should be followed!!! 1. LC or GLC transfer: If you already have an LC syringe, or just made some GLC from a clean jar of colonized grain, all it takes is 2-3cc of solution to get your jars going. Make sure you have clean culture first! 2. Grain transfer: The simplest way to inoculate your LC is to sprinkle in a couple colonized grains from a clean, fully colonized master grain jar. 3. Dowel spawn: I had made up a bunch of dowels of various species. Having never opened the sterile jars, I decided to inoculate a couple LC jars with colonized dowel plugs. The mycelium grew in a dense cluster off of the dowels and required vigorous swirling to break up the mass. This is only a viable option if you have sterilized dowels colonized, do not attempt with pasteurized dowels. Step 11: Sit back and watch you mycelium grow! Let it get a good foot-hold in the jar before you start swirling. When you swirl try to direct the mycelium across the tip of your downspout to slice it on the sharpened edges. Do this every couple of days until the myc has grow out enough to store the jars in the fridge. Keep an eye out for cloudiness of the liquid, or discolored spots growing on the sides of the jar or surface of the liquid. These are all signs of contaminated culture. Even without these signs, you should run a few test jars every now-and-then to make sure your culture is still clean. I usually G2G those clean test jars for a run of spawnings. Well folks, that's about it! Hope this helps to dispel the bad rep LC has gotten. Please try for yourselves and post your results here.
Edited by ghiajake (10/04/14 01:22 AM)
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EastBayRay

Registered: 06/06/13
Posts: 746
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Re: Liquid Culture De-Mystified: Ghia Style [Re: ghiajake]
#19541877 - 02/08/14 09:57 PM (9 years, 11 months ago) |
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Is your other handle "Morelman"?
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style [Re: EastBayRay]
#19541913 - 02/08/14 10:09 PM (9 years, 11 months ago) |
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Nope. No other handle.
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style [Re: ghiajake]
#19541982 - 02/08/14 10:30 PM (9 years, 11 months ago) |
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Note to mods: If this thread would be better suited in the "Mushroom Cultivation" forum, or any other forum, please feel free to move it.
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LustyLocks


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Re: Liquid Culture De-Mystified: Ghia Style [Re: ghiajake]
#19542092 - 02/08/14 10:58 PM (9 years, 11 months ago) |
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Wow. Very nice write up. Well done sir! 
To be honest. I cannot remember where I got my recipe. I looked at many to get ideas and ended up doing the same as you.
One thing I would like to add. When it comes time to add the ME, instead of just putting it in the pot I get two coffee filters and put my ME in there and then tie it off with a rubber band. That eliminates the whole pouring through a filter. You just have to wait for the ME to dissolve in the filter and then pour into jars. I've never had any sediment in my jars by doing it this way. I am pretty certain that I found that idea from monstermitch.
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Saint
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Re: Liquid Culture De-Mystified: Ghia Style [Re: ghiajake]
#19542106 - 02/08/14 11:02 PM (9 years, 11 months ago) |
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like your write up on making your LC I need to ask you about cloudiness of a grain LC does it need to be clear also. Before doing the LC I have a photo This is after grow in grain. 500ml of water use to pull. Was thinking I could expand with kero? In a new jar or inject sterile water/kero? not sure if i am on right track.
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style [Re: Saint]
#19542142 - 02/08/14 11:12 PM (9 years, 11 months ago) |
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I was doing it like that Clay, like you suggested before, but I was still getting the sediment. Weird, probably the filters I used.
Quote:
Saint said: like your write up on making your LC I need to ask you about cloudiness of a grain LC does it need to be clear also. Before doing the LC I have a photo This is after grow in grain. 500ml of water use to pull. Was thinking I could expand with kero? In a new jar or inject sterile water/kero? not sure if i am on right track.
GLC will usually be cloudy, but I don't know if I'd use what you have there. I suggest inoculating your LC straight from the syringe when you do your pull. Otherwise you're badly risking contamination. And you only wanna use a couple cc of GLC, more doesn't mean better. Your LC will be too cloudy to see through if you use anymore GLC than that.
Also, as I stated in the OP, don't even bother with karo or honey. It sucks, no matter what you've read. Trust me!
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Saint
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Re: Liquid Culture De-Mystified: Ghia Style [Re: ghiajake]
#19542174 - 02/08/14 11:23 PM (9 years, 11 months ago) |
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Quote:
ghiajake said:
GLC will usually be cloudy, but I don't know if I'd use what you have there. I suggest inoculating your LC straight from the syringe when you do your pull. Otherwise you're badly risking contamination. And you only wanna use a couple cc of GLC, more doesn't mean better. Your LC will be too cloudy to see through if you use anymore GLC than that.
Also, as I stated in the OP, don't even bother with karo or honey. It sucks, no matter what you've read. Trust me!
Thanks for leting me know about the grain looking cloudy i thought it would. Some one said not. I'll read some more to find what works the best for the shittake GLC I do like your write up here looks great.
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style [Re: Saint]
#19542199 - 02/08/14 11:32 PM (9 years, 11 months ago) |
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Just use the malt extract, it works for every thing. The first pic at the top is Shiitake LC made my way.
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Mrcloudy
Stranger than you.



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Re: Liquid Culture De-Mystified: Ghia Style [Re: ghiajake]
#19542283 - 02/09/14 12:29 AM (9 years, 11 months ago) |
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I don't understand the bad rep that liquid culture has gotten. I mean if your starting from spore, then yes it is a terrible idea. And I do understand that generally speaking you can't tell at a glance whether it is contaminated or not. But I have made several liquid cultures with several different species and have noticed that each one has a distinctive look and behaviour, and not all mycelium in LC looks the same. However if you spin it up every day it will mix it up and it will be a homogeneous blob which if you have some bad things in there they might be hard to distinguish.
I think it is similar to grain really. If you see anything that looks off, toss it. So far I have had no issues with properly made and inoculated LCs I have seen some spore LCs do some pretty disturbing things. If you start from spore, you're going to have a bad time. Inoculating with living mycelium is the way to go, I think a lot of people have issues because they start from spore. Not saying it is perfect as any lapse in sterile procedure and something could still get in but I did all my work in front of a impromptu "flowhood" and had no issues.
You should however always test your LCs on agar or a single jar before using it on a larger scale, just in case there are problems. I think a properly done LC can be a very useful tool in this hobby.
Excelent write up BTW!
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10 different Ganoderma species from across the USA AMU MrCloudys guide to North American GanodermaUpdated A rough guide to North American Ganoderma species, with an emphasis on the laccate species.
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style [Re: Mrcloudy]
#19542382 - 02/09/14 01:48 AM (9 years, 11 months ago) |
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Good to see another fan of LC.
I give each LC jar a test once a month at least. It is these test jars that I G2G from to expand my grain spawn
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Forrester
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Re: Liquid Culture De-Mystified: Ghia Style [Re: ghiajake]
#19542548 - 02/09/14 04:03 AM (9 years, 11 months ago) |
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I fucking love it, thanks for posting!
-------------------- Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here. ------------------- Have some medicinal mushrooms and want to get the most out of them? Try this double extraction method.
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style [Re: Forrester]
#19543116 - 02/09/14 10:01 AM (9 years, 11 months ago) |
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No prob Forrester, thanks.
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Saint
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Re: Liquid Culture De-Mystified: Ghia Style [Re: ghiajake]
#19543393 - 02/09/14 11:07 AM (9 years, 11 months ago) |
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This is a cube i started 3 days ago in a kero jar. It started out with the spore looking like black specks on the very bottom. Should of took a pic then. last night. This morning they formed in to this. I am going to try some of your incite to doing LC. I just like the grain LC and going to keep testing. Wish I could find more info on grain lc.
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style [Re: Saint]
#19543420 - 02/09/14 11:12 AM (9 years, 11 months ago) |
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Looks like good myc, but if you started from spores don't get your hopes up too soon.
What do you wanna know about GLC? There are some good teks on the subject floating around, but I can answer what you wanna know.
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cubenpete
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Re: Liquid Culture De-Mystified: Ghia Style [Re: ghiajake]
#19554116 - 02/11/14 07:29 PM (9 years, 11 months ago) |
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Very nice write-up, I like the straining suggestion, not sure why I havent done that before but I will now.
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RogerRabbit
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: ghiajake]
#19554257 - 02/11/14 08:02 PM (9 years, 11 months ago) |
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This thread was moved from Advanced Mycology.
Reason:
Quote:
ghiajake said: Anyone that has ever bought a culture syringe from a sponsor is using LC. If agar was SO much better than LC, then why don't the sponsors all sell wedges instead?
Because they're selling to noobs without a flowhood or skills to use agar.
Not an advanced mycology topic. Moved to cultivation. RR
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: RogerRabbit]
#19554307 - 02/11/14 08:14 PM (9 years, 11 months ago) |
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Quote:
Because they're selling to noobs without a flowhood or skills to use agar.
That may true, but it doesn't invalidate its usefulness. Merely casting LC aside once you increase you skills and equipment is a waste of a perfectly good tool.
Thanks for moving it to the appropriate forum RR.
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RogerRabbit
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: ghiajake]
#19554319 - 02/11/14 08:19 PM (9 years, 11 months ago) |
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Also, the reason new growers have trouble with water separating from the agar in petri dishes is due to improper storage. You're supposed to keep the petri dishes in an insulated cooler in the refrigerator to prevent the temperature swings from reaching the dishes. The insulated cooler does just that. RR
-------------------- Download Let's Grow Mushrooms semper in excretia sumus solim profundum variat "I've never had a failed experiment. I've only discovered 10,000 methods which do not work." Thomas Edison
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: RogerRabbit]
#19554816 - 02/11/14 10:15 PM (9 years, 11 months ago) |
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Thanks for that bit of info RR. I'll be sure to do that on the next batch to go in the fridge. I use a dorm fridge for my dishes. Any thing I could use for that, or do I need to get bigger fridge?
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MaJiK_420
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: ghiajake]
#19555116 - 02/11/14 11:44 PM (9 years, 11 months ago) |
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I tried and had one success and one failure with LC before
Kinda curios to try out this way, I have enough LME to last years with Agar, would that work for LC?
And I think I remember reading about no tilt jars before, but I can't remember why make it no tilt?
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: MaJiK_420]
#19555140 - 02/11/14 11:49 PM (9 years, 11 months ago) |
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Yes, lme is perfect. Use .6g of it for every 100mL of water, like I said in the OP. The no-tilt lids are much easier to extract from, you can use both hands on the syringe instead of holding the jar and one working the syringe. They aren't completely necessary, but I highly suggest them.
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x7x_x7x
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Re: Liquid Culture De-Mystified: Ghia Style [Re: ghiajake]
#19556839 - 02/12/14 11:22 AM (9 years, 11 months ago) |
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Quote:
Now, enough bragging, let's get down to business making some LC! Keep in mind, LC is meant for live culture transfers only!!! Trying to germinate spores in an LC solution will end in failure 9.9 times out of 10.
i germinated spores in LC many times and always work ok
-------------------- cultivando en la miseria SuctoSpore® Pictorial Tek
 x7x_x7x@shroomery.org carl_jung_in_lsd@yahoo.com koh samui and oak ridge are my favourite strains
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x7x_x7x
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Re: Liquid Culture De-Mystified: Ghia Style [Re: x7x_x7x]
#19556845 - 02/12/14 11:23 AM (9 years, 11 months ago) |
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-------------------- cultivando en la miseria SuctoSpore® Pictorial Tek
 x7x_x7x@shroomery.org carl_jung_in_lsd@yahoo.com koh samui and oak ridge are my favourite strains
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cronicr



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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: x7x_x7x]
#19556846 - 02/12/14 11:24 AM (9 years, 11 months ago) |
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insulated lunch boxes workk great i take mine out and leave it for the day before opening it
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  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
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iFung
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: cronicr]
#19556858 - 02/12/14 11:27 AM (9 years, 11 months ago) |
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I thought they sold spores as opposed to LC because spores are legal to sell while LC isn't? (at least among active strains)
--------------------
Get flushes like this from a half pint cake: Cake2Bowl Tek
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: iFung]
#19556909 - 02/12/14 11:46 AM (9 years, 11 months ago) |
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Quote:
i germinated spores in LC many times and always work ok
Awesome, I'll read the link in a minute. Doesn't concern me much since I don't grow actives, but I always love learning new shit.
Quote:
Insulated lunch boxes workk great
Thanks, I'll try that. Have to find one that'll fit right in there. Not too concerned, since I'm gonna probably have to get a bigger fridge anyways once it warms up enough I can't just use the shelf in my basement anymore.
Quote:
I thought they sold spores as opposed to LC because spores are legal to sell while LC isn't?(at least among active strains)
That is correct, but the OP was written to cover mostly edible/medicinal species. It works just as well with actives too.
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Saint
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: ghiajake]
#19566951 - 02/14/14 03:58 PM (9 years, 11 months ago) |
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i did find more info on my GLC thanks for looking Grain Liquid Culture (GLC) Tek
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: Saint]
#19566971 - 02/14/14 04:04 PM (9 years, 11 months ago) |
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I think I read that one too.
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kotter


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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: ghiajake]
#19579309 - 02/17/14 11:13 AM (9 years, 11 months ago) |
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Thanks for posting that write up about the value of liquid culture. One huge plus for LC in my world is the speed of preparation if I discover I want to make a clone immediately but don't have any appropriate media already made up. I can find a nice mushroom in the wild and make up some jars of liquid, have them pc'd, back to room temperature and have bits of fungal tissue in them within no more than a couple of hours. I've been using this for doing initial isolations for some months now and only going onto agar after I've got some clean looking growth going. Typically I move new growth onto agar at the same time I transfer the rest of the blobs onto some grain and a bit of it into a jar with some fresh LC. I try to keep a few extra on hand and they also seem to store better than agar. I've left a couple jars sitting around since October just to see how long they will last and neither has contaminated with anything yet. LC has proven itself to be a valuable approach for me this past year. Or at least, whatever the reason, my success rates for obtaining clean cultures from new clones has gone way up.
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blindingleaf
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: kotter]
#19579340 - 02/17/14 11:21 AM (9 years, 11 months ago) |
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nice write up!
i would add into the OP the option of putting an agar wedge into an LC to start it. works wonders as well. thats the only way i do it. the first time i used grains to noc up an lc, then when i started with agar, i moved to that.
jake, was wondering why u do not like honey or karo? i use honey, but am getting some LME in the mail soon. will substitute that for honey next time, but still curious.
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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Pastywhyte
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: blindingleaf]
#19579373 - 02/17/14 11:31 AM (9 years, 11 months ago) |
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I don't like LC for cubes at all but, I am a huge fan of it for p galindoi LC is an excellent choice for slow colonizing species. Just need to do it right. A large capacity syringe is a must IMO as you chance for contam increases every time you stick the SHIP. Larger syringe, less sticking. Little things like that are overlooked all to often with LC which IMO why it turns into a disaster.
My LC recipe is so simple. Take some grain soak water, run it through a coffee filter, dilute and use. Works fantastic IMO.
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: Pastywhyte]
#19579522 - 02/17/14 12:11 PM (9 years, 11 months ago) |
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Quote:
blindingleaf said: i would add into the OP the option of putting an agar wedge into an LC to start it.
Step 10, second sentence...
Quote:
If you have no problems with agar, then just noc up your LC with it if you like, but I think the following ways are less prone to contamination.
I just haven't had luck with using honey/karo. If it works for you keep using it, but I like the ME.
Quote:
Pastywhyte said: I Take some grain soak water, run it through a coffee filter, dilute and use. Works fantastic IMO.
I have done it like that in the past. When I was growing cubes I used the soak water from the straight grass seed I was using (mostly for agar, but made a couple LC). I had to severely dilute it to make it light enough to see through. Unfortunately, I hadn't built my flow hood yet and my jars got contamed. (I suck at agar in an SAB)
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: ghiajake]
#19583623 - 02/18/14 01:27 AM (9 years, 11 months ago) |
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Did up 6 more jars of LC tonight. I'll post pics once I see if there's any good growth.
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RandomFX
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: ghiajake]
#19593252 - 02/20/14 03:25 AM (9 years, 11 months ago) |
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way cool.
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RandomFX
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: RandomFX]
#19593273 - 02/20/14 03:33 AM (9 years, 11 months ago) |
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Quote:
Kotter said:I can find a nice mushroom in the wild and make up some jars of liquid, have them pc'd, back to room temperature and have bits of fungal tissue in them within no more than a couple of hours. I've been using this for doing initial isolations for some months now and only going onto agar after I've got some clean looking growth going.
you used Liquid Culture to isolate? how did you do that? Or do you mean just to initially hopefully weed out like debris or obvious mycelium growth to 'other things', then move to agar to actually isolate?
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kotter


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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: RandomFX]
#19596426 - 02/20/14 07:20 PM (9 years, 11 months ago) |
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Yes. I've found it really helpful for getting nice clones going of mostly wild mushrooms. I started doing this in an emergency pinch when finding a really nice hericium when I was leaving town within several hours. In that case I opened the PC while still it was still hot in front of the flow hood and let the airflow cool down the jar.
I'm essentially using liquid media for the initial isolation. If that growth does not come out clean I pitch it. I've learned the hard way that contaminants can lurk inside some of the tissues on hericium so don't trust them to be clean until after I've done two transfers with the last one being agar. I do that even if using agar for the initial isolation although lately I've been using part of the mycelium grown in liquid to inoculate some grain at the same time I'm inoculating agar. There has been very little incidence of contaminations so far.
I'm using a weak sugar solution (examples: 12 gm LME in a liter of distilled water or 4T Munton's malt extract in a liter or 19 grams light Agave nectar in a liter), cooking it 20 minutes in a half pint or pint mason jar with a filter disk lid, opening up a mushroom body in front of the hood, then teasing out a tiny piece that has never experienced the outer world until I tear it open. I usually make some short incisions around the outer portions of the target zone and then tease out bits of tissue with the tip of the scalpel or with a needle forceps. All of that is pretty normal stuff except maybe for my use of liquid instead of agar to start. I typically use a scalpel or a pair of needle forceps to fish out the wads of goo for transfers. In between transfers I drop the entire tool into a jar of 95% EtOH and then flame it until its dull cherry.
Hericium americanum in liquid malt extract and water

Local Hericium corolloides in agave nectar and water
Local Hericium erinaceus in spray-dried light malt extract and water
Local Pleurotus in agave nectar and water. The core was seriously dense and almost solid. It did not want to turn loose of the glass!
Local Trametes in agave nectar and water

A problem I encountered with this early on was mold riding inside of the Hericum tissues. I've come to believe its not always true but is almost always true when the tissue is soggy. I stopped taking samples from any soggy tissue (which means some specimens don't get cloned) and stopped trying to get branch or tip bits to grow and the incidence of mold plummeted to almost nothing. On a corolloides I feel fortunate if I can glean a single mm x a mm piece of clean tissue but one is all I need.
I'll add that the vast majority of anything I know came from comments on this forum. I know liquid does have a bad reputation in some people's opinion but I'd like to suggest that some of that might be not understanding some helpful things like never permitting a sample to float and not using compromised tissues. If any part of it has seen air other than from the flow hood or has touched another part of the mushroom during removal or been compromised by sog it seems likely it is going to require additional work to get it clean. (If liquid is exuded when the sample is being taken its often a waste of time - this is common for erinaceus.) If the tissue is clean and only moist like normal healthy mushroom tissue a tiny bit is all it has been taking. The base of Hericiums has been the easiest place to find that sort of sample but any inner part that is thick enough to be torn open seems to work too. Obviously I try to get this completed as soon as possible after finding a mushroom. I now try to keep a few jars of liquid made up and on hand at all times just in case something interesting pops up.
The way I'm doing it is not amplifying like Ghia Jake's sweet write up above but is just another application of the same sort of media.
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: kotter]
#19596515 - 02/20/14 07:45 PM (9 years, 11 months ago) |
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Dude, thanks for posting this! It's awesome. I will be trying this with some wild specimens this year for sure! Have you ever included an autoclavable anti-bacterial agent in LC cloning solution? Do you dip the wild specimen in H2O2 before tearing and taking a sample? This is a great way conserve dishes and reduce waste stream.
This is what I like, other LC believers coming out of the closet and promoting useful applications of this tool.
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RandomFX
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: ghiajake]
#19596834 - 02/20/14 09:08 PM (9 years, 11 months ago) |
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Quote:
All of that is pretty normal stuff except maybe for my use of liquid instead of agar to start.
ah ok...so I am getting the impression you really use it just to start a culture, not really to isolate. you just get the culture going, hopefully from a clean sample since you are taking the sample from the interior and then you actually isolate on agar later, am I understanding correctly? I have no problem with LC's, as was stated earlier, they are a tool and a great one for mass inoculation since they can easily spread and thin the live culture out more so then a solid on agar, clearly. I would be surprised if professionals did not use them for bulk inoculation of spawn, and in fact most of us do in reality use them when we put agar pucks in deionized water for long term storage. that is clearly really a type of liquid culture, and in many cases the strain can be stored according to papers I've read, I haven't had them that long to attest to it myself for a 20 yr storage, at ambient temperature none the less. I personally say screw slants...but well I do not know. some species may not like that type of LTS. I did read where a huge culture bank did go completely to DDH2O storage in like 1994 or something, and they obviously know what they are doing, so I am fairly safe in the theory of it working pretty well, I think.
Edited by RandomFX (02/20/14 09:13 PM)
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PussyFart
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: RandomFX]
#19596857 - 02/20/14 09:13 PM (9 years, 11 months ago) |
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Quote:
RandomFX said: I would be surprised if professionals did not use them for bulk inoculation of spawn.
A lot of them don't.
A lot of them G2G, which is safer and quicker in most cases.
--------------------
THIS HOBBY IS NOT FOR THE IMPATIENT! PLEASE BE PATIENT, DON'T BE A PATIENT! A Tale of 10 Isolates, GT Cluster Clone Monotubs, RR's Let's Grow Mushrooms DVD, SGFC(Shotgun Fruiting Chamber), Monotub Tek, Damion5050's Coir Tek, TL's Tek List, Frank's Tek List, EvilMushroom666's Pasteurization Tek, How It Should & Shouldn't Look - NEW CULTIVATORS GUIDE *** *** AFGHAN KUSH GROW LOG *** ***
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: RandomFX]
#19596879 - 02/20/14 09:19 PM (9 years, 11 months ago) |
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Quote:
I am getting the impression you really use it just to start a culture, not really to isolate. you just get the culture going, hopefully from a clean sample since you are taking the sample from the interior and then you actually isolate on agar later, am I understanding correctly?
Technical it is a cloning step, but further isolation may be needed to produce a mono culture. I'm sure it's mono by the second dish though, am I right kotter? Have you been able to do this with any mychorrizal species?
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Pastywhyte
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: kotter]
#19596969 - 02/20/14 09:37 PM (9 years, 11 months ago) |
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Quote:
kotter said: A problem I encountered with this early on was mold riding inside of the Hericum tissues. I've come to believe its not always true but is almost always true when the tissue is soggy.
I had mold issues trying to clone wild H coralloides as well. Fortunately my buddy (who found the location where we got them) took a spore print, and so I have managed to get a good ms culture going (with the help of some tetra agar) and now have a few small ms subs going. Hopefully I can see fruits and clone from those
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kotter


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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: Pastywhyte]
#19597264 - 02/20/14 10:41 PM (9 years, 11 months ago) |
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Good call! I've been using the word 'isolate' completely wrongly. I think cloning is the right word. My goal is not actually to isolate a strain - I'm just trying to acquire mushrooms that I can propagate and enjoy using wild ones as my starting point. It would seem like its already a monoculture at the point of taking that small bit of mushroom tissue? If I was starting from spores I'd need to go through several plates to have a monoculture since I'd need to isolate a sector first and then be sure it was true.
Thanks all for straightening out my bad use of language!
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PussyFart
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: kotter]
#19597297 - 02/20/14 10:47 PM (9 years, 11 months ago) |
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Quote:
kotter said: It would seem like its already a monoculture at the point of taking that small bit of mushroom tissue?
--------------------
THIS HOBBY IS NOT FOR THE IMPATIENT! PLEASE BE PATIENT, DON'T BE A PATIENT! A Tale of 10 Isolates, GT Cluster Clone Monotubs, RR's Let's Grow Mushrooms DVD, SGFC(Shotgun Fruiting Chamber), Monotub Tek, Damion5050's Coir Tek, TL's Tek List, Frank's Tek List, EvilMushroom666's Pasteurization Tek, How It Should & Shouldn't Look - NEW CULTIVATORS GUIDE *** *** AFGHAN KUSH GROW LOG *** ***
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: kotter]
#19597306 - 02/20/14 10:48 PM (9 years, 11 months ago) |
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I like the idea of cloning the best MS fruit, then going from there. Isolating a strain before you've ever see how it fruits seems a little backwards.
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kotter


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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: kotter]
#19597318 - 02/20/14 10:50 PM (9 years, 11 months ago) |
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And no, I don't use an antibiotic.
I tried using commercial antibiotic agar when I was battling to get my first clean corolloides cultures in 2011. It seemed to be of help when I was using a laborious series of transfers from agar to agar. This is not laborious but choice of tissue is probably crucial. Liquid seems pretty unforgiving when it contaminates.
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: kotter]
#19597486 - 02/20/14 11:44 PM (9 years, 11 months ago) |
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Yeah, two of the 4 LC's I just did are contamed. The other 4 are going strong and I'll be testing them by next week.
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RandomFX
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: ghiajake]
#19598120 - 02/21/14 04:25 AM (9 years, 11 months ago) |
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Quote:
A lot of them don't. A lot of them G2G, which is safer and quicker in most cases.
ya, guess the introduction of added water gives a place for contaminants to creep in so it is a added layer of risk. and thinning the myc out more surely adds more risk also, since that is often when myc is the weakest when it is thin and just starting out and spreading, I would think.
Do those using LC, use LC for spawn inoculation, then spawn to substrate, or right to substrate inoculation? (Not counting using a jar to test the purity or agar/etc.)
Quote:
Yeah, two of the 4 LC's I just did are contamed. The other 4 are going strong and I'll be testing them by next week.
If a experienced person such as yourself is having problems of 50% contamination, I'm thinking the case for LC is diminishing. This is I am sure the biggest problem with LC, is it's spread so thin and homogeneous inside that if it gets contaminated, there isn't much you can do. where as agar...you can easily isolate away from it simply by cutting one or a few good pieces away, if it out races it, in even the smallest of locations. I still think LC is cool though and surely useful making a quart of the stuff and using a spray bottle as you walk through the woods one day has a appealing quality to me over sprinkling agar or grain, and surely sticks to trees better. but it seems G2G is safer and more solid.
Edited by RandomFX (02/21/14 04:36 AM)
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blindingleaf
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: RandomFX]
#19598363 - 02/21/14 07:04 AM (9 years, 11 months ago) |
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the only time i actually "need" an lc is if i do filter bags of grain b/c i have no hood. so i inject thru the bag then cover it back up in an sab. u could wedge a bag I'm sure, but at that point u might as well G2G if u have to open the bag all the way anyway in an SAB. and i only use an lc if it was nocced up with an agar wedge. anyways, my two cents.
and even tho stamets has sections about LC in GGMM (and TMC to a lesser extent) Pussyfart is right, just watch any video on u tube after typing "mushroom growing" or "mushroom farm" and u will see them (edible mushroom growing operations) ALL g2g and then spawn to bags with grain. its generally agar wedge to grain master, grain master to X amount of 2nd gen quarts, then those quarts to filter bags, then fruit in/thru/with those bags. although I'm basing this off of u tube videos, so this speculation/observation could be inaccurate.
maybe amanita virosa could chime in here, as he works for spawn production company (right?), so he may know best what the current commercial practices are for edibles.
jake, how did u know ur lc was contaminated? color, growth, something else?
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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Stargaze
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Re: Liquid Culture De-Mystified: Ghia Style [Re: ghiajake]
#19598446 - 02/21/14 07:32 AM (9 years, 11 months ago) |
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Quote:
ghiajake said: De-vilifying an extremely useful and easy tool: by GhiaJake
If agar was SO much better than LC, then why don't the sponsors all sell wedges instead?
Wouldn't that be illegal?
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Pastywhyte
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Re: Liquid Culture De-Mystified: Ghia Style [Re: Stargaze]
#19598519 - 02/21/14 08:05 AM (9 years, 11 months ago) |
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Not for edibles or medicinal mushrooms. Fact is that you can't buy active LC either, you buy spore syringes.
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blindingleaf
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Re: Liquid Culture De-Mystified: Ghia Style [Re: Pastywhyte]
#19598575 - 02/21/14 08:31 AM (9 years, 11 months ago) |
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 I'm pretty sure sponsors sell lc's b/c they are easier for beginners to use, which i would assume is what they are counting on for their main source of income? if u look at the prices of an lc versus a slant (the other option from vendors in regards to edibles) there is at least a $!0 difference in price, slants being higher. this makes sense tho, a company with a good flow hood and professional personnel can make a clean quart of lc with an agar wedge in the same time it takes to make a few slants. the lc quart will grow out enough to make many many lc syringes, making it cheaper to produce, while coincidentally (maybe on their part?) being easier to work with for most beginners. i buy the lc's from edible vendors, but i still work at an hourly job making hoagies, so i can't afford slants ATM. i just noc up plates, then store those. same culture, different price. actually, now that i think about it, why would I or anyone buy the slants if u are already working with agar? just make ur own and save $. only benefit i can guess with slants is long term viability, which includes shipping/receiving, but if they are a good vendor, they will replace a non viable LC culture that died in transit at god's speed (they did in my case anyway).
even better, look at the market place here, or just troll the forums and grow logs. ask around, offer a trade, u never know what kinds of stuff u can get, for free, or thru trades. who needs money or bit coins when u have an amazing culture to exchange?
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style [Re: blindingleaf]
#19598810 - 02/21/14 09:46 AM (9 years, 11 months ago) |
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Quote:
If a experienced person such as yourself is having problems of 50% contamination, I'm thinking the case for LC is diminishing. This is I am sure the biggest problem with LC, is it's spread so thin and homogeneous inside that if it gets contaminated, there isn't much you can do.
I did make a typo, it was 2 out of 6, with 4 still good. To be fair, I did noc all 6 with grains from jars I already G2G'd other grains with, so that's most likely the cause. If I had used agar I'd be throwing away money (dishes) right now instead of just dumping them and trying again.
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Mrcloudy
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Re: Liquid Culture De-Mystified: Ghia Style [Re: ghiajake]
#19598874 - 02/21/14 10:03 AM (9 years, 11 months ago) |
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It seems likely that inoculating with grains was the vector for contamination.It might be a good idea to keep a couple of agar plates aside simply for LC inocculation. 2 out of 6 is not all too bad, especially when like you said you just dump out and start over, little bit of nutrients and a little bit of water and your good to go.
Its good to see people actively working with LCs instead of giving up because of contamination issues. I think a lot of the issues can be solved with relatively little effort.
--------------------
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PirateSwazey



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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: Mrcloudy]
#19599191 - 02/21/14 11:10 AM (9 years, 11 months ago) |
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Nice write up Jake 
Where are you getting your stainless tubing to make your stirrers?
Don't give up on your agar work, if I can do it you can brudda
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: PirateSwazey]
#19599360 - 02/21/14 11:56 AM (9 years, 11 months ago) |
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Quote:
Its good to see people actively working with LCs instead of giving up because of contamination issues.
I can guarantee that 100% of my contaminations are due to my lack judgement. I should have noc'd the LCs first, then G2G'd after, not G2G then LC a week later. But, in my defense, I was probably stoned at the time. 
Quote:
Where are you getting your stainless tubing to make your stirrers?
I ordered it on Ebay from Zoro Tools, I buy it in 6' lengths (best bargin). Decent price, plus it comes in a 7' long 3" cardboard tube. Who doesn't need one of those!?! I think I'm gonna cut it in 4 sections and stuff them with sawdust and spawn, make some cardboard logs. That cardboard is rather dense. Y'all think I should drill some small holes in them, or let the mycelium just colonize the cardboard and fruit on its own? I could use a plant based sealant on the outside to combat contaminants until the mycelium fully colonizes the cardboard and eats it's way through the sealant from the inside. What do y'all think?
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Pastywhyte
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Re: Liquid Culture De-Mystified: Ghia Style [Re: ghiajake]
#19599374 - 02/21/14 11:59 AM (9 years, 11 months ago) |
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Quote:
ghiajake said: If I had used agar I'd be throwing away money (dishes) right now instead of just dumping them and trying again.
Sounds like you just need a good no pour tek that uses cheap and reusable containers
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style [Re: Pastywhyte]
#19599468 - 02/21/14 12:29 PM (9 years, 11 months ago) |
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I'll probably pick up some regular mouth clear walled 1/4 pint jelly jars as soon as I can find them. Only ones around here are the squat wide mouth, and the "dimpled" wall regs. The "whatman" filter lids should be fine for agar jars, right? I like the idea of being able to cut wedges, or use a syringe and sterile water to extract the myc. And not have to buy dishes.
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YOmamaPr0
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Re: Liquid Culture De-Mystified: Ghia Style [Re: ghiajake]
#19599498 - 02/21/14 12:38 PM (9 years, 11 months ago) |
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PirateSwazey



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Re: Liquid Culture De-Mystified: Ghia Style [Re: ghiajake]
#19599631 - 02/21/14 01:14 PM (9 years, 11 months ago) |
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Quote:
ghiajake said: The "whatman" filter lids should be fine for agar jars, right? I like the idea of being able to cut wedges, or use a syringe and sterile water to extract the myc. And not have to buy dishes.
Sounds badass
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RandomFX
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Re: Liquid Culture De-Mystified: Ghia Style [Re: ghiajake]
#19599907 - 02/21/14 02:29 PM (9 years, 11 months ago) |
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Quote:
ghiajake said: I did make a typo, it was 2 out of 6, with 4 still good. To be fair, I did noc all 6 with grains from jars I already G2G'd other grains with, so that's most likely the cause. If I had used agar I'd be throwing away money (dishes) right now instead of just dumping them and trying again.
Fair enough, but that is still 33% but then again it is water and myc...not like a major expense. As far as the dishes. use glass petri's. I do, and do not throw any of them away, and I have two sleeves of disposables that I haven't opened since. cynmar.com sells glass petri's for .80 a piece in packs of 10. for 15mm x 75mm I believe.
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style [Re: RandomFX]
#19600109 - 02/21/14 03:18 PM (9 years, 11 months ago) |
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Glass petris are awesome, but I can't do a liquid extraction like with the jars. I will probably pick some up eventually, just because they're cool. I picked up 5x 2ml glass syringes for that very same reason. Gonna order a 100mL one eventually too.
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kotter


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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: ghiajake]
#19664651 - 03/07/14 06:40 PM (9 years, 10 months ago) |
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Maybe LC does have limited usefulness but it enabled this clone to enter my life:
On grain: two jars of erinaceus; a jar of corolloides is on the right.

erinaceus

No idea why I could not get the above two not to rotate. I unchecked the pertinent box to no avail.
This is my very first lion's mane fruit from a clone I made!
Several more bags are close behind (just as I'm leaving town for a week)


The original fruiting body it came from:
Edited by kotter (03/07/14 06:41 PM)
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blindingleaf
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: kotter]
#19664661 - 03/07/14 06:42 PM (9 years, 10 months ago) |
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nice pics kotter!
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: blindingleaf]
#19664829 - 03/07/14 07:34 PM (9 years, 10 months ago) |
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Thanks! I'm still pretty low on the learning curve but have sure become obsessed with Hericiums. I've halfway filled a room with bags on racks. I'll post more images when I get back home.
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invitro


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Re: Liquid Culture De-Mystified: Ghia Style [Re: x7x_x7x]
#19664855 - 03/07/14 07:39 PM (9 years, 10 months ago) |
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Quote:
x7x_x7x said: read SuctoSpore® Pictorial Tek
I read this tek, but I don't really see the advantage of using a 3 way stop cock. I've never seen one up-close but my main concern is how do you sterilize the part of the stop cock that receives the needle? If your familiar with Tl's no tilt lc tek, this seems to offer the same functionality and the rtv blob is easy to sterilize with alcohol.
http://www.shroomery.org/forums/showflat.php/Number/14186384
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ghiajake
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Re: Liquid Culture De-Mystified: Ghia Style [Re: invitro]
#19664886 - 03/07/14 07:47 PM (9 years, 10 months ago) |
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Nice looking culture and fruits kotter!
invitro: I briefly debated using female luer-lock tips for my LC lids, but not with a check valve like that. I was going to use caps for them instead. Didn't use them because I already had the parts for the no-tilts I made.
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invitro


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Re: Liquid Culture De-Mystified: Ghia Style [Re: ghiajake]
#19664956 - 03/07/14 08:13 PM (9 years, 10 months ago) |
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What I really like about the rtv blobs is that you can wipe alcohol on them wet, with a small square of sponge, then ring out the same sponge and kind of dab it 'dry' Leaving what is probably a very thin film of alcohol which probably evaporates nearly instantly. The evaporation of alcohol from a contam spore is actually what kills it, bursting it from the inside (I read that somewhere). A 3 way port or a luer lock would just kind of stay wet. I'm not sure if having it stay wet is actually a disadvantage but it seems like it might be. Someone who is more up on science would have to chime in there.
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Re: Liquid Culture De-Mystified: Ghia Style [Re: invitro]
#19665115 - 03/07/14 09:04 PM (9 years, 10 months ago) |
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Silicone injection ports are always a risk. The best you can do is wipe them with alcohol which is only a sanitizer not a sterilizer so you always have a small chance of pushing contams through no matter what. Same reason why no one wipes with alcohol after flaming it's pointless. Alcohol than flame, You can't flame a silicone injection port so you're always trying your luck and every single time you use a LC you have to test it.
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Re: Liquid Culture De-Mystified: Ghia Style [Re: bodhisatta]
#19665140 - 03/07/14 09:12 PM (9 years, 10 months ago) |
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That's why I use as large a syringe as possible and crack the lid when inoculating the cut down those vectors. Though even a 60 cc syringe may test clean on the first couple jars, but after doing 10-12 more it might have picked up a contam for the last one or two. That's why you must treat LC like a grenade. It has great power, but can go off in yer face if your not careful.
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YOmamaPr0
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Re: Liquid Culture De-Mystified: Ghia Style [Re: Pastywhyte]
#19665185 - 03/07/14 09:26 PM (9 years, 10 months ago) |
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Like a grenade. Metaphorically beautiful. *technically a simile but it didn't sound as good.
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Forrester
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Re: Liquid Culture De-Mystified: Ghia Style [Re: bodhisatta]
#19665199 - 03/07/14 09:31 PM (9 years, 10 months ago) |
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Quote:
bodhisatta said: Silicone injection ports are always a risk. The best you can do is wipe them with alcohol which is only a sanitizer not a sterilizer so you always have a small chance of pushing contams through no matter what. Same reason why no one wipes with alcohol after flaming it's pointless. Alcohol than flame, You can't flame a silicone injection port so you're always trying your luck and every single time you use a LC you have to test it.
Very good point - unless, you just don't remove the foil after PC, until cooled and ready to inject. Foil off, BAM! inject. Pretty sterile-like
-------------------- Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here. ------------------- Have some medicinal mushrooms and want to get the most out of them? Try this double extraction method.
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Re: Liquid Culture De-Mystified: Ghia Style [Re: bodhisatta]
#19665227 - 03/07/14 09:36 PM (9 years, 10 months ago) |
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So I am new at working with Hericium, not to detract from the conversation about Ghias liquid culture. But is that whispy branching common in hereciums? Several months ago I found a dried sad looking mushroom fruiting from pine and identified it as Hericium americanum. Since this was the only one I could find that had been collected from conifers before. By some bizzare miracle, perhaps due to the mother mushroom being immature I was able to clone and clean up a culture without the nightmare that I have read about with cloning lions manes. I had zero bacterial issues, but a few fungal contams.
I have been apprehensive about growing it out because I have suspected it was a contaminant. The culture has been sitting in my fridge this whole time because I have a terrible habit of doing that. But I guess what I want to know is does this look like Herecium mycelium? It has a similar whispy branchlike growth thing going on. I have been meaning to grow it out for testing but havent gotten around to it yet and didn't want to waste time trying to grow something that wasn't what I was after.

Again sorry, I don't mean to detract from this thread, I have just been wondering about my culture for a while now and that jar pic there reminded me I still have this thing hanging out in there.
--------------------
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Re: Liquid Culture De-Mystified: Ghia Style [Re: Mrcloudy]
#19665248 - 03/07/14 09:42 PM (9 years, 10 months ago) |
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Detract away my friend.
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invitro


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Re: Liquid Culture De-Mystified: Ghia Style [Re: Forrester]
#19665304 - 03/07/14 09:55 PM (9 years, 10 months ago) |
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Quote:
Forrester said:
Quote:
bodhisatta said: Silicone injection ports are always a risk. The best you can do is wipe them with alcohol which is only a sanitizer not a sterilizer so you always have a small chance of pushing contams through no matter what. Same reason why no one wipes with alcohol after flaming it's pointless. Alcohol than flame, You can't flame a silicone injection port so you're always trying your luck and every single time you use a LC you have to test it.
Very good point - unless, you just don't remove the foil after PC, until cooled and ready to inject. Foil off, BAM! inject. Pretty sterile-like 
Yea that's a good idea, plus if your needle goes into the ship when the needle was red-hot 2 seconds ago, anything that might get "pushed through" is probably going to get burned to death on the way in. You wouldn't think the silicone could handle that kind of heat but the red rtv silicone can take it. You might singe it a little though.
Edited by invitro (03/07/14 09:56 PM)
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bodhisatta 
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Re: Liquid Culture De-Mystified: Ghia Style [Re: invitro]
#19665336 - 03/07/14 10:01 PM (9 years, 10 months ago) |
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even regular silicone will handle it, Silicone caulking is all 100% silicone and can easily take the red hot needle, the RTV just means it cures a bit differently (room temperature vulcanizing IE could cure underwater and without fresh air) you can cure it in the PC as soon as it forms a skin instead of waiting 24+ hours. The needle cools quite a bit very quickly and it has a low thermal mass, temperature doesn't kill bacteria temperature+time kills bacteria so it's very easy for the bacteria to survive the hot needle poke if the port is dirty, or the alcohol failed to kill something. It's nothing to get crazy worried about but if things go wrong, it's one of the possibilities. This is why most of us don't bother with LC, LC is good for vendors to sell edible/medicinal tried and tested monoculture rather than having you grow from spores and get average genetics, it doesn't mean it's a great way to inoculate and store inoculant, especially for actives like cubes.
Edited by Trusted cuItivator (03/07/14 10:03 PM)
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invitro


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Re: Liquid Culture De-Mystified: Ghia Style [Re: bodhisatta]
#19665371 - 03/07/14 10:12 PM (9 years, 10 months ago) |
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Do you have a reference that could expand on this idea of heat + time kills bacteria. I can understand maybe if the temperature increase was gradual you might say that, like starting up a pot of boiling water or a pressure cooker. But lets say there is some bacteria on the ship and the still very hot needle hits it, that kind of instant temperature change would undoubtedly decrease the amount of time needed to kill the bacteria dramatically don't you think? Kind of an analogy would be glass jars, you can heat them up in a pc over 30 minutes and they won't break, but try applying rapid heat, like a flame, to the jar, it'll shatter.
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bodhisatta 
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Re: Liquid Culture De-Mystified: Ghia Style [Re: invitro]
#19665429 - 03/07/14 10:26 PM (9 years, 10 months ago) |
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needles are hollow bacteria are microscopic the area around the needle will have very slow heat transfer through the silicone and then the jabbing could dislodge or rub them against the inside of the needle which has cooled down who knows but peoples LC contaminates even when made from agar wedges all the time having been clean for a few weeks.
direct flame will oxidase/incinerate anything the flame is nearly 2000F from butane if I remember right but you can see how quickly your needle ceases to stay red hot, it's probably a long shot but I don't discount it considering the fate of most LC after repeated stabbings.
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invitro


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Re: Liquid Culture De-Mystified: Ghia Style [Re: bodhisatta]
#19665446 - 03/07/14 10:32 PM (9 years, 10 months ago) |
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Even though the needle is not red anymore when it hits the port, it's still plenty hot. When I accidentally bump the plunger on the syringe and a drop comes into the needle, the water vaporizes and steam shoots out of the needle tip.
Innoculating a lc with an agar wedge is not smart, I will agree with that. Doing it the way I said seems to be pretty effective IME.
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bodhisatta 
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Re: Liquid Culture De-Mystified: Ghia Style [Re: invitro]
#19665459 - 03/07/14 10:34 PM (9 years, 10 months ago) |
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Quote:
invitro said: Even though the needle is not red anymore when it hits the port, it's still plenty hot. When I accidentally bump the plunger on the syringe and a drop comes into the needle, the water vaporizes and steam shoots out of the needle tip.
Innoculating a lc with an agar wedge is not smart, I will agree with that. Doing it the way I said seems to be pretty effective IME.
Inoculating a lc with an agar wedge is not smart*
Agar wedge is the best way, but even then they can still contam and it's likely from the needle poking. Also I suspect a lot of people inadvertently cool their needle by plunger pressing as they stick the injection port
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invitro


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Re: Liquid Culture De-Mystified: Ghia Style [Re: bodhisatta]
#19665464 - 03/07/14 10:38 PM (9 years, 10 months ago) |
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It depends on your environment, if you have a high spore count in the air, I wouldn't try it even with a sab. With a flowhood or a relatively clean work area sure.
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Re: Liquid Culture De-Mystified: Ghia Style [Re: invitro]
#19665481 - 03/07/14 10:41 PM (9 years, 10 months ago) |
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with a high spore count a SAB has to be just as effective it's all or nothing, the SAB works by not having air currents and the drag of particles encountering the still air. If you're using your SAB the right way it wont really matter. But, If you have a high spore load and use a Flow hood you're getting 99.97% filtration some stuff will make it through, that's why you work fast even in front of a flow hood(they're not sterile TEK saviors you still need excellent sterile TEK and rehearsed agility) , and it's also why the opening the petri and letting the FH blow on it isn't a good test to see if you built your FH right because even a properly built one will fail that test if you leave the dish open long enough.
Edited by Trusted cuItivator (03/07/14 10:42 PM)
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invitro


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Re: Liquid Culture De-Mystified: Ghia Style [Re: bodhisatta]
#19665502 - 03/07/14 10:47 PM (9 years, 10 months ago) |
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What I'm saying is a SAB isn't effective enough (to get away with agar wedges in an lc) with a high spore count, because you have to put your gloves through the box, there is always some air current it's juut minimal and minimal is too much in a very high spore load situation IME.
Edited by invitro (03/07/14 10:48 PM)
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bodhisatta 
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Re: Liquid Culture De-Mystified: Ghia Style [Re: invitro]
#19665508 - 03/07/14 10:49 PM (9 years, 10 months ago) |
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IME you wipe your gloves with iso inside of the SAB, it's what field mycologists use in the middle of a jungle for wild samples with success, Imagine the spore count in a humid jungle.
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Mrcloudy
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Re: Liquid Culture De-Mystified: Ghia Style [Re: bodhisatta]
#19665523 - 03/07/14 10:51 PM (9 years, 10 months ago) |
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Yeah I have not had any issues inoculating LC with agar wedge In my opinion its just about the best way to go about it, treat it like a grain jar. Open it up drop the wedge in quickly then cover it back up working as quickly as you can. I admit that opening the jar is nerve racking, anything that is a potential vector of contamination makes me nervous. But no more so than doing a grain jar.
I have not yet had a contamination doing it this way. I have had some funky looking LCs though, but that was due to the variety of ways each species deals with being submerged.
--------------------
10 different Ganoderma species from across the USA AMU MrCloudys guide to North American GanodermaUpdated A rough guide to North American Ganoderma species, with an emphasis on the laccate species.
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Re: Liquid Culture De-Mystified: Ghia Style [Re: bodhisatta]
#19665529 - 03/07/14 10:52 PM (9 years, 10 months ago) |
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all i've ever done with lc's is wedges in my sab, still air is still air, spritz water around and get to work
--------------------
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Re: Liquid Culture De-Mystified: Ghia Style [Re: cronicr]
#19665787 - 03/08/14 12:01 AM (9 years, 10 months ago) |
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The long term storage by distilled water method, is pretty much a LC. And by pretty much I mean it is period.
I have to believe it has to be quite viable for so many major people and institutions to use it, and there is one major mycology collections group named with over 3,000 recorded specimens at the writing of the article that named them, that switched over totally to distilled water for long term storage in 1994. although I am currently doing it and testing it specifically per species I have access to, and it has been used for decades, and the specimens have been revived after decades as well. I won't say what it works with or does not specifically, I haven't tested it yet, but there's some pretty darn convincing argument for it. LC's obviously work, and work well from agar. All my species are LTS in distilled water, and not refrigerated, and only in small 13mm x 100 mm culture tubes that I get for .48 cents ea. and I am willing to bet, in the next week when I revive those cultures, which holds about 12 cc's ea. I'll easy be able to grow from less than a drop of culture, which means if the standard of 20 drops/ml is used, then a 12 ml vial of LTS mycelium can potentially revive the strain 240 times, easy. Not bad imo.
http://www.hardydiagnostics.com/articles/Saving-fungal-Cultures-Sterile-Water-By-Harris.pdf
http://www.scielo.br/scielo.php?pid=s0365-05962005000700004&script=sci_arttext&tlng=en "CONCLUSIONS
Based on the experiment, it can be concluded that:
1. Strain preservation by the distilled water method has made it possible, in a short period of time, to prove the viability and sporulation capacity of the strains submitted to this study.
2. Maintenance technique in sealed flasks containing distilled water has proven to be efficable for preserving the characteristics of sporulation of fillamentous fungi of medical interest.
3. Conservation of fungi in flasks with distilled water has easy handling, storage and transportation, besides being more economic the traditional method of successive samplings"
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blindingleaf
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Re: Liquid Culture De-Mystified: Ghia Style [Re: RandomFX]
#19666069 - 03/08/14 01:43 AM (9 years, 10 months ago) |
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yea I'm with cron i only do agar to lc in an SAB. its same as agar to agar or agar to grain. the lc will start faster if u drop in a couple wedges or at least scratch up a larger wedge then drop it in, so when u shake, little parts of the myc come off the wedge and start their own colonies. i also use no RTV port or ge port, easier to shake rigorously. i only fill up the needle ONCE, when lc is two weeks in. test it on agar. if its clean, u have made a pre made sterile stringe, similar to what an edible vendor would send u. the trick is to get as much myc in ONE suck up as u can. that way u only open the lc jar twice, once to inoculate with a wedge, twice to suck up the lc with the needle. failure is still an option in any case.
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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tombosley8
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Re: Liquid Culture De-Mystified: Ghia Style [Re: blindingleaf]
#19666420 - 03/08/14 05:36 AM (9 years, 10 months ago) |
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wow this is the info i've been looking for i've always feared the lc and the stigma around it but if I could make a syringe with one little wedge that'd be awesome. Can you store this at room temp or fridge for long durations?. or am i not seeing something. Yes syringe inoc seems very easy compared to agar but what are the draw backs. Just more chance of contam I guess because of the TWO times you need to open jar but seems better than exposing agar for every jar to inoc and maybe this is utilizing the agar for more quantity?
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blindingleaf
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Re: Liquid Culture De-Mystified: Ghia Style [Re: tombosley8]
#19666440 - 03/08/14 05:47 AM (9 years, 10 months ago) |
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it does have a lot of stigma. mainly b/c its an older flash in the pan style of cultivation, at least here in the forums. most ppl, when u read those old teks, are using either spore solution or clone tissue. u cannot see a contam on either of those before inoculation ur LC solution. if u start ur LC with agar, u can see a contam (most likely). i still do agar to grain 95% of the time. i ONLY use lc if I'm doing filter bags of grains, b/c i don't trust myself in an SAB unfolding the bag to dump qt of grain in. some people drink less coffee than me tho, so probably have steadier hands and g2g with filter bags are no problem for them. wish i could, g2g is faster than lc.
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Pastywhyte
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Re: Liquid Culture De-Mystified: Ghia Style [Re: blindingleaf]
#19666727 - 03/08/14 08:40 AM (9 years, 10 months ago) |
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I gotta agree that the best way to inoculate a LC is a clean agar wedge. MS syringes are almost never clean and using one to inoculate a LC is a huge contam vector.
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RandomFX
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Re: Liquid Culture De-Mystified: Ghia Style [Re: tombosley8]
#19668180 - 03/08/14 03:39 PM (9 years, 10 months ago) |
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Quote:
tombosley8 said: wow this is the info i've been looking for i've always feared the lc and the stigma around it but if I could make a syringe with one little wedge that'd be awesome. Can you store this at room temp or fridge for long durations?. or am i not seeing something. Yes syringe inoc seems very easy compared to agar but what are the draw backs. Just more chance of contam I guess because of the TWO times you need to open jar but seems better than exposing agar for every jar to inoc and maybe this is utilizing the agar for more quantity?
Read the urls I provided. it is why I provided them. There are more layers of contamination with liquid culture. first off I think when hyphae is young it is far more susceptible to contamination and has less defense to fight it off (Now it's defense may even be only something as simple as the hartig net which may hang up contamination particles for all I know and not allow them conditions to fruit or thrive. or as has been mentioned the mycelium may lay down either chemicals of it's own, or remove all the yummy nutrients and leave the area barren in a burn the fields type of defense too....I do not know, haven't followed up on it. but young hyphae is evidently more susceptible and in liquid culture, if it has nutrients that move around, plainly one could see how after being put into a spawn or other thing it could easily be susceptible also. Where as a agar wedge, is developed. and then it spreads from sort of a 'base of operations and safety', and is able to consistently reinforce itself and thus maybe fight its way through. furthermore, if you are using a nutrient rich liquid, you are going to be feeding the nasties also...they love wet and nutrient rich. agar doesn't spread the moisture love all over the place. furthermore, the concentration of the mycelium is far higher on agar pure volume of a top layer of mycelium, and it's nutrients that it is using stay with it. a small square of mycelium hartig mat can have numerous layers upon layers of cells and hyphae strands. and many more supporting the ones that are there. it is true, in a liquid culture you can have tons of cells to duplicate, but on the top of colonized agar it is far more dense with way more cells. The liquid culture will probably have more 'strands' ie more of the strings will be cut up and smaller, but that only leaves them more open to attack, not less, I would think.
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EastBayRay

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Re: Liquid Culture De-Mystified: Ghia Style [Re: bodhisatta]
#19669417 - 03/08/14 08:44 PM (9 years, 10 months ago) |
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Quote:
bodhisatta said: Silicone injection ports are always a risk. The best you can do is wipe them with alcohol which is only a sanitizer not a sterilizer so you always have a small chance of pushing contams through no matter what. Same reason why no one wipes with alcohol after flaming it's pointless. Alcohol than flame, You can't flame a silicone injection port so you're always trying your luck and every single time you use a LC you have to test it.
That's why you apply enough High Temp RTV silicone so that you can push a RED HOT needle through it and when you pull the needle out it will seal completely.
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Re: Liquid Culture De-Mystified: Ghia Style [Re: EastBayRay]
#19669462 - 03/08/14 08:51 PM (9 years, 10 months ago) |
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all silicone is high temp. RTV just means it will cure without air "room temp vulcanizing" The Red RTV us for automotive use and has some additives for corrosion resistance but it's got the same heat properties that any 100% silicone has including oven mits made from silicone etc...
besides I seriously doubt the needle has enough thermal mass and time to kill everything just from being red hot, the red hot kills anything on the needle.
Edited by Trusted cuItivator (03/08/14 08:52 PM)
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EastBayRay

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Re: Liquid Culture De-Mystified: Ghia Style [Re: bodhisatta]
#19680400 - 03/11/14 11:10 AM (9 years, 10 months ago) |
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Quote:
bodhisatta said: all silicone is high temp. RTV just means it will cure without air "room temp vulcanizing" The Red RTV us for automotive use and has some additives for corrosion resistance but it's got the same heat properties that any 100% silicone has including oven mits made from silicone etc...
besides I seriously doubt the needle has enough thermal mass and time to kill everything just from being red hot, the red hot kills anything on the needle.
Well, it's worked for me for years. Try it for yourself using proper sterile technique.
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bodhisatta 
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Re: Liquid Culture De-Mystified: Ghia Style [Re: EastBayRay]
#19680444 - 03/11/14 11:28 AM (9 years, 10 months ago) |
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I have great sterile technique, that's why I do things the right way with agar and grain transfers rather than bullshit needle pokes. I got past that stuff after like a month of dicking around with that noob bs. 
Those techniques like SHIPs etc.. make this hobby foolproof so that anyone who can flush a toilet can have a moderate to good level of success with a low amount of failures.
Edited by Trusted cuItivator (03/11/14 11:29 AM)
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EastBayRay

Registered: 06/06/13
Posts: 746
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Re: Liquid Culture De-Mystified: Ghia Style [Re: bodhisatta]
#19683135 - 03/11/14 09:30 PM (9 years, 10 months ago) |
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Quote:
bodhisatta said: I have great sterile technique, that's why I do things the right way with agar and grain transfers rather than bullshit needle pokes. I got past that stuff after like a month of dicking around with that noob bs. 
Those techniques like SHIPs etc.. make this hobby foolproof so that anyone who can flush a toilet can have a moderate to good level of success with a low amount of failures.
I wasn't even insinuating you use improper technique or that you aren't experienced. Don't get all butt hurt, Jesus.
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kotter


Registered: 01/15/11
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Re: Liquid Culture De-Mystified: Ghia Style [Re: Mrcloudy]
#19712539 - 03/18/14 10:30 AM (9 years, 10 months ago) |
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Mrcloudy: Try transfering some of that culture to a thick layer of agar or some grain inside of a half pint mason jar with a filter patch and forget about it for a while in a cool location. Americanum tends to fruit small and fairly fast in that situation. It would be an easy way to tell if its right. For me, whispy branching has been common & Hericium species cultures enjoy being stored in a refrigerator -- even when forgotten for more than a year. Not too long ago I found a corolloides I'd somehow overlooked from late in 2012 and its doing great on grain and more recently on sawdust.
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Mrcloudy
Stranger than you.



Registered: 10/01/13
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Re: Liquid Culture De-Mystified: Ghia Style [Re: kotter]
#19712946 - 03/18/14 12:13 PM (9 years, 10 months ago) |
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Quote:
kotter said: Mrcloudy: Try transfering some of that culture to a thick layer of agar or some grain inside of a half pint mason jar with a filter patch and forget about it for a while in a cool location. Americanum tends to fruit small and fairly fast in that situation. It would be an easy way to tell if its right. For me, whispy branching has been common & Hericium species cultures enjoy being stored in a refrigerator -- even when forgotten for more than a year. Not too long ago I found a corolloides I'd somehow overlooked from late in 2012 and its doing great on grain and more recently on sawdust.
That is encouraging , thank you. I will definitely try it out.
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10 different Ganoderma species from across the USA AMU MrCloudys guide to North American GanodermaUpdated A rough guide to North American Ganoderma species, with an emphasis on the laccate species.
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kotter


Registered: 01/15/11
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Re: Liquid Culture De-Mystified: Ghia Style [Re: Mrcloudy]
#19714197 - 03/18/14 04:15 PM (9 years, 10 months ago) |
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I wish I had a better image but this one was chopped up a bit for propagation. Despite that it will probably give a better idea than my words. All three hericium species growing here seem to be able to do this. A bit of tin foil for sake of spore collection is blocking much of the fruiting body that formed. What is growing on agar in this jar was originally transferred there from liquid culture growing from a wild fragment.



This is one I have some ID questions about I had posted earlier elsewhere here (#18933228). As soon as I have those spores I can complete that ID question appropriately. It grows up to resemble americanum more than erinaceus but if so should not be growing wild here. I thought I'd let my americanum escape but tracking back through my records showed that to be wrong.
Edited by kotter (03/18/14 04:28 PM)
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Saint
Pope



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Re: Liquid Culture De-Mystified: Ghia Style [Re: kotter]
#19758886 - 03/28/14 06:07 AM (9 years, 9 months ago) |
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I know I will find my answer to question the more I read. Still going to ask about Temp to use when making liquid culture or expanding liquid culture in small 150cc jar's Is there a max temp to use. My temp for these shiitake LC been from 86-100 degrees. seen to be ok only 2 days.
    I stir by hand as much as possible. the drake specks is from grain Turkey baster is to big. Using 3ml glass storage vials 
I did have one issue I made some 1000 cc jars growth was great pulled most of the mycaleum off used for sawdust blocks so i know the LC was good. But I did make up sterile water and added 700cc to the jars. I all so used honey to make them. Added 20cc of sterile honey. thinking it would help the mycelium to grow. I think i killed my jars.IT looks like brown crap/dust not fluffy like normal. Still learning the hard way. CAN SOMEONE ADD WATER/FOOD TO A LC JAR?
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blindingleaf
blue collar underworld



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Re: Liquid Culture De-Mystified: Ghia Style [Re: Saint]
#19758958 - 03/28/14 07:11 AM (9 years, 9 months ago) |
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86-100!?!?! wow there captain sunbeam, lets bring it down! 
shoot for no more than 80, i'd say less, like 78. if u want faster growth, use less nutrients. (I'm using 1.5% sugars now, i think the norm is 4%)
the last part of ur post...not sure what u mean? seems like u attempted either a GLC (or sawdust LC?) . thats a good route (I never did it, do not trust it, but seems to work for ppl) but the nutrients are sorta kinda already in there, and come out when u suck the myc water back out (some nutrients are extracted from the short soak btw injection and sucking back up). if u added honey to it, u would have needed to sterilize it. even then, not the best idea. what u should do, If I'm understanding ur goal correctly, is do a grain LC (shoot sterile water into sterile grain jar 100% colonized) then suck it back out, and shoot THAT into a pre made, pre sterilized LC. u do not even have to go that far, that GLC is ready to use as it comes out of the grain jar, but if u wanna expand it, thats the route to take.
jake can chime in here, as he is way more experienced with edibles than i am, but i ONLY start LC's with an agar wedge. i kno glc seems like a good idea, but i still don't trust it like i do with agar. even an agar LC i will test before using.
also, cannot tell by the pics, but are u using an SAB or a hood? that will increase ur success rate a thousand fold, with any part of this hobby.
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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Saint
Pope



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Re: Liquid Culture De-Mystified: Ghia Style [Re: blindingleaf]
#19759046 - 03/28/14 07:58 AM (9 years, 9 months ago) |
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shoot for no more than 80, i'd say less, like 78. if u want faster growth, use less nutrients. (I'm using 1.5% sugars now, i think the norm is 4%)
Thank's for the temp advise I was just playing around to see what temp change would do. normally i do have them sitting in cobert around 80 degrees. Just this time was trying something new with the LC And had to ask what effect temp does have. other them contaminants grow better at higher temps.
2) the last part of ur post...not sure what u mean? seems like u attempted either a GLC (or sawdust LC?) .
No, not a glc or sawdust LC. I Have akot of them bags already IT's a straight LC I ll take a photo of what happen when i added water and honey {yes all sterilize}. added a photo in minute be right back
I have to remove some photos it looks like before i can upload new ones as i could not upload. thanks for let me know about the 1.5 % i ll try that
Edited by Saint (03/30/14 03:18 PM)
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invitro


Registered: 05/03/13
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Re: Liquid Culture De-Mystified: Ghia Style [Re: Saint]
#19759606 - 03/28/14 11:49 AM (9 years, 9 months ago) |
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What variety of shitake do you have? I think there are cold weather and warm weather types.
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Saint
Pope



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Re: Liquid Culture De-Mystified: Ghia Style [Re: invitro]
#19769146 - 03/30/14 03:14 PM (9 years, 9 months ago) |
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I am not even sure which one i have growing right now i have like 5 different ones I growing now waiting to see the fruit to see which is going to be a few months yet. KMH,DCL,96,SBW
Right now trying to figure out how to do this spore growth with Petri dish. And its just pissing me off with these cubes first black mold/green mold now I am brown contam's When i get one good dish i will be happy. The shiitake has be very easy to keep clean nothing get contam so I know its the spore syringe's
Forgot to say i started this on 01/01/2014 I never did this before. But i do have almost 100 blocks made up all ready. IT's been very easy you just need right equipment.
Edited by Saint (03/30/14 03:26 PM)
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blindingleaf
blue collar underworld



Registered: 07/19/13
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Re: Liquid Culture De-Mystified: Ghia Style [Re: Saint]
#19769201 - 03/30/14 03:25 PM (9 years, 9 months ago) |
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that sucks man, i had an unlucky cube print like that once. do u have any current grows u can clone from?
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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Saint
Pope



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Re: Liquid Culture De-Mystified: Ghia Style [Re: blindingleaf]
#19769229 - 03/30/14 03:31 PM (9 years, 9 months ago) |
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Quote:
blindingleaf said: that sucks man, i had an unlucky cube print like that once. do u have any current grows u can clone from?
Tell me about it. at least I learning how to do everything first hand. I got ahold of vendor on here and he replaced my first synge with 3 new ones. So he was very good to me. i am sure I will get one to work. Just the time is really the only thing I am out. No cubes as of yet But i am learning about spores.
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blindingleaf
blue collar underworld



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Re: Liquid Culture De-Mystified: Ghia Style [Re: Saint]
#19769240 - 03/30/14 03:33 PM (9 years, 9 months ago) |
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nice! I'm sure at least one of those will be clean. i have only had one bad syringe. u can also hit the marketplace and do trades. maybe if u have an edible culture u can trade that for a cube print or two.
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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Saint
Pope



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Re: Liquid Culture De-Mystified: Ghia Style [Re: blindingleaf]
#19769277 - 03/30/14 03:41 PM (9 years, 9 months ago) |
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Quote:
blindingleaf said: nice! I'm sure at least one of those will be clean. i have only had one bad syringe. u can also hit the marketplace and do trades. maybe if u have an edible culture u can trade that for a cube print or two.
Good idea I am just not ready yet I have only bout 70 master 3 ml vials made up so far. trying to keep my self supplied with grain spawn is my main idea right now. i want to make at least 100 blocks per month so i can sell them at framers market with the mushrooms I grow. Its my goal to have a part time job from a hobby.
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blindingleaf
blue collar underworld



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Re: Liquid Culture De-Mystified: Ghia Style [Re: Saint]
#19769292 - 03/30/14 03:44 PM (9 years, 9 months ago) |
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100 blocks!! jeez. i live in a place thats too hot for cold species in the summer like shiitaki. i'd need to run the AC window unit a lot, even then, it still is about 80 minimum in my 3rd floor apt. i have 2 shiitaki bags going now, pop corning. I'm hoping they beat the warm weather that is surely coming. just got our first big spring rain all day yesterday, continuing on right now.
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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cronicr



Registered: 08/07/11
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Re: Liquid Culture De-Mystified: Ghia Style [Re: blindingleaf]
#19769316 - 03/30/14 03:47 PM (9 years, 9 months ago) |
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yeah i kinda missed my shitake window lazy stoner season
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  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
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Saint
Pope



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Re: Liquid Culture De-Mystified: Ghia Style [Re: blindingleaf]
#19769396 - 03/30/14 04:00 PM (9 years, 9 months ago) |
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Well I don't worry about elc its in my rent for a apt on the 16th floor. I have the green house set up in front of a 12 foot window with a AC unit right on the same wall. Have a walk in closet i am using to store the blocks in the dark ruff 78 degrees so i am just waiting to start see them all browning at one time. I am not worried about cost and will sell blocks for $15 at the market just enough to cover my cost. With some extra But if I sell a bag of grown your going to pay for them! Retired in 2004 so its just a hobby to me.
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blindingleaf
blue collar underworld



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Re: Liquid Culture De-Mystified: Ghia Style [Re: Saint]
#19769599 - 03/30/14 04:43 PM (9 years, 9 months ago) |
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oh u mean ur selling kits? i thought u meant actual fruits
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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madLyfe
Wonderer



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Re: Liquid Culture De-Mystified: Ghia Style [Re: blindingleaf]
#20122494 - 06/13/14 12:38 AM (9 years, 7 months ago) |
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i am using the TL's not tilt tek, but was wondering what other things could be used to break up the LC other than rocks?
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ghiajake
Myco-Viking


Registered: 01/10/13
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Re: Liquid Culture De-Mystified: Ghia Style [Re: madLyfe]
#20655885 - 10/04/14 12:47 AM (9 years, 3 months ago) |
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Quote:
madLyfe said: i am using the TL's not tilt tek, but was wondering what other things could be used to break up the LC other than rocks?
Best thing is a magnetic stirrer.
Just noc'd up a new PM LC a couple days ago. This time I used grass clipping/reishi tea (boiled hard for an hour) as the nutrient base. Been using this as my agar nutrient base with good success so I figured I'd try it in LC too. I'll post a pic in a couple days.
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eatyualive
Eat's You Alive :)



Registered: 08/17/01
Posts: 19,026
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Re: Liquid Culture De-Mystified: Ghia Style [Re: ghiajake]
#20662205 - 10/05/14 03:59 PM (9 years, 3 months ago) |
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beautiful work!
makes me want to bust out my ancient stir plate and stir bar.
Edited by eatyualive (10/05/14 04:00 PM)
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