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Offlineghiajake
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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)

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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Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: kotter]
    #19596969 - 02/20/14 09:37 PM (9 years, 11 months ago)

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 :cool:


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Offlinekotter
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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)

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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OfflinePussyFart
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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)

Quote:

kotter said:
It would seem like its already a monoculture at the point of taking that small bit of mushroom tissue?



:justno:


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Offlineghiajake
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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)

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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Offlinekotter
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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)

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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Offlineghiajake
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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)

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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OfflineRandomFX
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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)

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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Invisibleblindingleaf
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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)

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?


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OfflineStargaze
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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)

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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InvisiblePastywhyteMDiscord
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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)

Not for edibles or medicinal mushrooms.  Fact is that you can't buy active LC either, you buy spore syringes.


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Invisibleblindingleaf
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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)

:whathesaid:
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?


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Offlineghiajake
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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)

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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OfflineMrcloudy
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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)

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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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 (moved) [Re: Mrcloudy]
    #19599191 - 02/21/14 11:10 AM (9 years, 11 months ago)

Nice write up Jake :thumbup:

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 :grin:


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Offlineghiajake
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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)

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. :wink:

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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Re: Liquid Culture De-Mystified: Ghia Style [Re: ghiajake]
    #19599374 - 02/21/14 11:59 AM (9 years, 11 months ago)

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 :wink:


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Offlineghiajake
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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)

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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OfflineYOmamaPr0
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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)

:goodluck:


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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)

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 :super:


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