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numnum59
Pro-Am Mycologist

Registered: 12/07/11
Posts: 1,189
Last seen: 1 year, 5 months
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liquid culture inoculation
#15720179 - 01/26/12 10:16 AM (12 years, 11 months ago) |
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Is it possible to use too much lc when inoculating. I've used 1cc on each inoculation site before but what about 1.5
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oysterizer


Registered: 12/20/11
Posts: 183
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Re: liquid culture inoculation [Re: numnum59]
#15720243 - 01/26/12 10:31 AM (12 years, 11 months ago) |
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Using to much lc can throw off the moisture content. 1.5 cc should be fine though. Try it and if it goes slower it got to wet.
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Icesyn
Trust My Words


Registered: 05/19/11
Posts: 1,880
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Re: liquid culture inoculation [Re: oysterizer]
#15720277 - 01/26/12 10:40 AM (12 years, 11 months ago) |
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It is understandable that you believe more LC would speed things up.
As the last dude said, moisture is where it could go wrong. Allot of people who prep grain fail to remember that before loading them up, they should be soaked and then dried correctly. Otherwise, you'll face the most common slap in the face, bacteria.
Remember, you don't need much LC to get it started. To much can ruin it and waste LC if the LC is even good at that.
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Shroom_Goon
Just Some Guy

Registered: 08/11/11
Posts: 475
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Re: liquid culture inoculation [Re: Icesyn]
#15720333 - 01/26/12 10:57 AM (12 years, 11 months ago) |
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I recently used 5cc of LC on each grain jar. Haha I had too much of the shit and was wondering how fast it would colonize. Took about 2 weeks, including the shake, for full colonization. I was stoked.
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Icesyn
Trust My Words


Registered: 05/19/11
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Re: liquid culture inoculation [Re: Shroom_Goon]
#15720362 - 01/26/12 11:05 AM (12 years, 11 months ago) |
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You were stoked, while your grains were soaked.
I'm not a gambling man. Gradz on a good LC though.
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numnum59
Pro-Am Mycologist

Registered: 12/07/11
Posts: 1,189
Last seen: 1 year, 5 months
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Re: liquid culture inoculation [Re: Icesyn]
#15723198 - 01/26/12 11:27 PM (12 years, 11 months ago) |
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one of my liquid cultures froze and the other two got really cold, do u think they are still viable?
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oysterizer


Registered: 12/20/11
Posts: 183
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Re: liquid culture inoculation [Re: numnum59]
#15723892 - 01/27/12 06:06 AM (12 years, 11 months ago) |
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Most cultures do not freeze well, but the ones that did not freeze should be fine. Its a good idea to do a test jar anyway.
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Galaxie500
Rat Bastard



Registered: 11/08/11
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Re: liquid culture inoculation [Re: oysterizer]
#15724215 - 01/27/12 09:13 AM (12 years, 11 months ago) |
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I keep my LC's covered with foil in the fridge. It slows 'em down (but they're definitely still growing, or my eyes are tricking me). I don't even let it warm up when I use it, straight from the fridge to the jar. I use TranscendingLife's ELME LC's and they work fantastic, DESPITE the fact I used too much ELME AND they caramelized.
As far as amounts...
I use A LOT (now, I didn't to start with). I usually used about 4cc's making BRF cakes, and I use 10cc's in my WBS jars now. My WBS colonizes in 8 days, like clockwork. It definitely makes a difference in time; I inoculate down the sides of the jars and then distribute however much is left after that all over the top of the grain, so instead of a small splotch germinating, a huge portion germinates all at once. I usually hit ~20% for the shake in about 3 days. I don't use the textbook recipe and methodology, however. I make my grains/cakes intentionally drier than specified to compensate for the extra moisture I'm adding. With BRF cakes it was fairly easy, I would essentially just subtract out at least 4cc's from the recipe. It's a little less exacting for WBS, I spread it all out over a table and let it sit for a couple hours versus letting it just drain out of the colander.
It's always a nice surprise waking up a couple days after inoculation to discover that half the sides of the jars are colonized. There definitely was a difference when I made/make BRF cakes, but there's a HUGE difference in the WBS jars.
Edited by Galaxie500 (01/27/12 09:16 AM)
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numnum59
Pro-Am Mycologist

Registered: 12/07/11
Posts: 1,189
Last seen: 1 year, 5 months
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Re: liquid culture inoculation [Re: Galaxie500]
#15725536 - 01/27/12 03:34 PM (12 years, 11 months ago) |
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Thanks galaxy
And what's the best way to make a test jar
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Infinitys Minute
a universe inside each moment


Registered: 11/02/11
Posts: 1,757
Loc: Zion
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x
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All my posts are completely fictional and are for entertainment purposes only, images I use, post or reference on this account are sourced from the internet and are not my own.
Edited by Infinitys Minute (10/04/16 01:27 AM)
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rony-pony
Stranger
Registered: 03/17/11
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Re: liquid culture inoculation [Re: numnum59]
#15726743 - 01/27/12 07:50 PM (12 years, 11 months ago) |
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Morning all,
I was wondering if anyone could point me in the direction of a step by step tek for making LC from colonized rye grain? Is it correct to assume that LC's go a lot further? - how long do they last... as in how long can you take from one LC (Time-wise I mean, does the LC regenerate?) Also is it commonly agreed that straw logs are a safer bet than laundry baskets? Cobweb seems to be a recurring problem. How can this be prevented?
Will the same LC tek work for spore syringe?
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oysterizer


Registered: 12/20/11
Posts: 183
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Re: liquid culture inoculation [Re: rony-pony] 1
#15728312 - 01/28/12 06:42 AM (12 years, 11 months ago) |
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To make lc from colonized grain, simply inject jar with sterile water and shake the hell out of it. Then suck out the liquid and your ready to noc up jars. I try to use lc made like this within a week. You could squirt a little into some premade lc and grow more. My lc's that are grown in malt water will last about a year in the fridge, but many contaminate after pulling from them a couple times. I like to make several jars and plan to use all of one jar at a time. Contamination can be very hard to see in lc. The myc will regenerate in lc as long as there is still nutrition to feed it,which there wont be if pulled from a jar of grain with sterile water. Lc can stretch a long way, you dont need much as long as you have patience. In my opinion patience is key to growing mushrooms. Cobweb can be prevented with enough fae and lower humidity, or sprayed with hydrogen peroxide.
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rony-pony
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Re: liquid culture inoculation [Re: oysterizer]
#15729254 - 01/28/12 01:00 PM (12 years, 11 months ago) |
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That's great, thanks.
To prevent cobweb and other contams do you think using a hair dryer round to dry the edges of the straw log would help if there is no H202 at hand, until H2O2 is bought Two weeks in and the mycellium hasn't taken full control, perhaps a not enough grain was used.
I agree with you, patience and attention to details go a very long way.
An expert is someone who has made all the mistake already! You really do learn from your mistakes
Thanks
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PinsWellWithOthers
Thread Derailer



Registered: 10/15/10
Posts: 1,834
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Re: liquid culture inoculation [Re: rony-pony]
#15729304 - 01/28/12 01:18 PM (12 years, 11 months ago) |
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Edited by PinsWellWithOthers (03/06/12 05:44 PM)
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oysterizer


Registered: 12/20/11
Posts: 183
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Re: liquid culture inoculation [Re: rony-pony]
#15730234 - 01/28/12 05:13 PM (12 years, 11 months ago) |
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Quote:
rony-pony said: That's great, thanks.
To prevent cobweb and other contams do you think using a hair dryer round to dry the edges of the straw log would help if there is no H202 at hand, until H2O2 is bought Two weeks in and the mycellium hasn't taken full control, perhaps a not enough grain was used.
I agree with you, patience and attention to details go a very long way.
An expert is someone who has made all the mistake already! You really do learn from your mistakes
Thanks
Not sure about the hair dryer, havent tried that before. H2o2 just melts cobweb right before your eyes, so i never tried anything else after that.
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gizmo1



Registered: 06/15/11
Posts: 3,831
Loc: FREEDOM
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Re: liquid culture inoculation [Re: numnum59]
#15731008 - 01/28/12 08:07 PM (12 years, 11 months ago) |
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Quote:
numnum59 said: Thanks galaxy
And what's the best way to make a test jar
This is probably gonna be your best bet bro. http://www.shroomery.org/forums/showflat.php/Number/15713099#15713099
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rony-pony
Stranger
Registered: 03/17/11
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Last seen: 12 years, 8 months
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Re: liquid culture inoculation [Re: rony-pony]
#15731701 - 01/28/12 11:03 PM (12 years, 11 months ago) |
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Where would you typically get H202, ie is it something commonly found in shops/chemists?, and what % is best to use?
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oysterizer


Registered: 12/20/11
Posts: 183
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Re: liquid culture inoculation [Re: rony-pony]
#15732556 - 01/29/12 06:36 AM (12 years, 11 months ago) |
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Any pharmacy or store that sells band aids. In with the first aid by rubbing alcohol. Its sold at 3% and that is a good strength for cobweb. Usually costs about a dollar. It is sold at a higher concentrations at hydro shops but that stuff is stronger usually 30 %
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rony-pony
Stranger
Registered: 03/17/11
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Re: liquid culture inoculation [Re: oysterizer]
#15732671 - 01/29/12 08:05 AM (12 years, 11 months ago) |
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Excellent, thanks very much, I'll dander to the shop now and see what I can find, does it come in liquid form? If so how is best to apply evenly? Thanks for your patience in answering my questions, I just want to try and keep mistakes to a minimum
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oysterizer


Registered: 12/20/11
Posts: 183
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Re: liquid culture inoculation [Re: rony-pony]
#15732759 - 01/29/12 08:45 AM (12 years, 11 months ago) |
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It comes in liquid form in a brown bottle, so as not to let any light in. Light breaks it down into water and oxygen. Put some in a spray bottle and mist affected areas. You dont want to use it anymore than is necessary to control it as it slows down mushroom myc growth as well. I have completely soak sub in h2o2 in the past when the cobweb was really bad, and it did eventually recover and fruit but it would have been faster to throw it out and make another.
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rony-pony
Stranger
Registered: 03/17/11
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Re: liquid culture inoculation [Re: rony-pony]
#15732797 - 01/29/12 09:02 AM (12 years, 11 months ago) |
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I went to three different pharmacies, none had any, one just didn't have any in stock which is good news for future but for now it leaves me in trouble - I picked up cream peroxide 6%, a hair product but I'm not too sure if this will have the desired effect. Obviously if there's H2O2 in it that's great but it will also have other things in it. What do you think? give it a bash? I'm going away for a few days first thing in the morning so would rather do something now before it's too late
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oysterizer


Registered: 12/20/11
Posts: 183
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Re: liquid culture inoculation [Re: rony-pony]
#15732865 - 01/29/12 09:30 AM (12 years, 11 months ago) |
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Hmm, as far as i know benzoyl peroxide is used for hair bleaching/dye. Benzoyl peroxide is usually prepared by treating hydrogen peroxide with benzoyl chloride, and i have no idea how that will affect the sub in the long run. It leaves behind benzoic acid as it breaks down,which its salts are a food preservative with antifungal properties. Although it inhibits mold yeast and bacteria i havent found anything that says it inhibits mushroom myc. It could be an interesting experiment, but risky. If your situation is bad it may be worth a try diluted to 2% or 3%. Dilute some with water and try on a small test area.
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rony-pony
Stranger
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Re: liquid culture inoculation [Re: rony-pony]
#15732928 - 01/29/12 09:53 AM (12 years, 11 months ago) |
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I diluted the solution - added the same volume again in water - and gave a light spray to the vertical edges of the basket, and a couple of drops on the top. Situation isn't bad but if untreated it would go bad, just a couple of little wisps had appeared. Prevention is better than cure and all that... hopefully it will work out fine, either way, an experiment and chance to learn!
When I mentioned the hair dryer earlier, my thinking was that if the edges were dry then they'd be a lot less likely to host contams, like a kind of dry buffer area an inch thick all round, leaving the core to keep growing, it could also be worth a go next time. Or move on to the logs though from if they are less prone to contams.
With regards to LCs, can you effectively go on infinitely once up and running. ie LC from colonised grain(1), innoc more grain(2), LC from fruit of first grain, innoc more grain(3), LC from fruit of second set of grain, innoc.... and so on. What are common pitfalls?
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oysterizer


Registered: 12/20/11
Posts: 183
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Re: liquid culture inoculation [Re: rony-pony]
#15733146 - 01/29/12 11:19 AM (12 years, 11 months ago) |
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For the most part fresh air and control of humidity is the best way to prevent the cobweb from ever happening. If you create dry areas with hair dryer your myc wont colonize well, and potentially harmful mold can grow even if moisture content is low. Best to treat or cut away infected areas. How are you providing fresh air exchange?
As cultures grow they essentially age. The more times a culture has been grown out and cloned the older and weaker it will become. You can do this for a while but eventually it will be to weak to colonize or fruit well. If you search senescence you should find out more. The closer the culture is to the original spores the younger it is. Agar in slants or petri dishes are good for making a master culture from a good clone, then using those to grow out new lc. You can use lc as a master culture and squirt a little into a fresh jar to become the next master, but eventually it will decline.It is advisable to learn to take spore prints of whatever your growing,so you can start over if cultures are lost to senescence or contam. One print should have enough spore to go the rest of your life. Contams are near impossible to see in liquid unless it goes completely cloudy. You always want to inoculate some test spawn first just to verify that there arent any unwelcome guests so you dont waste time and material growing a dozen jars of mold. I prefer agar to lc because contams are visible and you can rescue you cultures by transferring to a clean dish. One dish can make more spawn(or lc) than you know what to do with. Its pretty easy to do even with just a glovebox.
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rony-pony
Stranger
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Re: liquid culture inoculation [Re: rony-pony]
#15733263 - 01/29/12 11:49 AM (12 years, 11 months ago) |
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I've been fanning the fruiting chamber 4-5 times per day, or basically every time I walk past it. I have been doing spore prints already and have had good success, the last one I done was huge and got me 30 syringes, each with visible spores, however each set of jars from these became contaminated, so I've been trying to learn about varying methods to see which works best for me. That's not to say I'll not do spore prints because of one failed batch, just trying to learn more and see which ways minimise the chance of contams. That way I am not relying soley on one method and can have a better all round understanding. What is the best way to store a print?
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PinsWellWithOthers
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Registered: 10/15/10
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Re: liquid culture inoculation [Re: rony-pony]
#15733687 - 01/29/12 01:29 PM (12 years, 11 months ago) |
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Edited by PinsWellWithOthers (03/06/12 05:32 PM)
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rony-pony
Stranger
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Cheers for the print tip.
Unfortunatley a bit of green mould took hold along with cobweb, it appeared in the last couple of days. The straw log was done three weeks ago today, so I guess there wasn't enough grain to completely colonise it quick enough.
Oh well, onwards and upwards, we learn from our mistakes
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