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 Cyber
Ash
 
Registered: 06/14/04
Posts: 1,041
Loc: Dearborn Michigan
Last seen: 29 days, 19 hours
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 Culture storage
#8328704 - 04/25/08 09:51 PM (6 months, 23 days ago) |
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Ok, this is a few questions
First, how long will a spore syringe stay viable in the fridge? One of the Species, that I grew out and printed, seems to not want to germinate. So I still have the original spore syringe in the fridge from 2004. What is the chance it is still good?
Second, how long will live cultures keep in the fridge? I just put live slants of Hypsizygus marmoreus, Pleurotus eryngii, Pleurotus ostreatus, Lentinula edodes, and Pleurotus Citrinopileatus in the fridge. How long will they keep and/or should I make plates from them every once in a while and refresh the slants?
Third, Is there a good place to look for growing parameters of the edibles? In what conditions do Lentinula edodes do best? etc. etc.
Thanks
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 fastfred
Registered: 05/17/04
Posts: 4,360
Loc: Dark side of the moon
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Re: Culture storage [Re: Cyber]
#8329491 - 04/26/08 03:48 AM (6 months, 23 days ago) |
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Syringe in the fridge should be good for several years. Your 2004 one is probably still good.
I had a culture last nearly a year in the fridge. It was a little overgrown when I put it in there and it was pretty weak when I used it. It was only about 50% viable using about 2cc per jar. But if it had been stored before getting as overgrown it probably would have survived better.
Everything should be transferred once in awhile. I'd say you'll want to revive and transfer every 6 months or so. RR probably has a better idea on timelines.
-FF
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tripsis
biophile-misanthropist-doofer
Registered: 04/13/08
Posts: 4,123
Loc: Gondwana
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Re: Culture storage [Re: fastfred]
#8329781 - 04/26/08 08:12 AM (6 months, 23 days ago) |
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Since we depend on an abundance of functioning ecosystems to cleanse our water, enrich our soil and manufacture the very air we breathe, biodiversity is clearly not an inheritance to be discarded carelessly - Edward O. Wilson 1992
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x7x_x7x
urban farmer
Registered: 05/05/07
Posts: 679
Last seen: 1 hour, 6 minutes
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Re: Culture storage [Re: tripsis]
#8329930 - 04/26/08 09:44 AM (6 months, 23 days ago) |
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http://www.shroomery.org/8509/Resurrecting-a-Better-Method-for-Long-Term-Storage-of-Mushroom-Cultures
"I came upon an article written by Michael D.Graham,a microbiologist at ATCC (1) that described the storage of yeast in sterile distilled water. What a brilliant idea! If that method stores yeast, It should work well on gourmet mushroom cultures, too. It's easy to do, very space efficient, allows the cultures to be stored at room temperature, and maintains thier viability for years. I contacted the author, he indicated that edible fungi stores even better than yeast, and you can store the spores as well as the hyphae often for decades!"
my fucking God what a data!
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transhumanism + primitivism=magic mushrooms
SuctoSporeŽ
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RogerRabbit
Bans for Pleasure
Registered: 03/26/03
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Loc: USA Mountain Northwest
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Re: Culture storage [Re: x7x_x7x]
#8330037 - 04/26/08 10:29 AM (6 months, 23 days ago) |
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I put a hardwood stick in each slant, and they easily last 2 to 3 years under refrigeration. If the mycelium on the agar dies, it will still revive from the wood. I use medical tongue depressors.
If you don't use wood, make fresh slants every 6 to 8 months.
RR
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Cyber
Ash
Registered: 06/14/04
Posts: 1,041
Loc: Dearborn Michigan
Last seen: 29 days, 19 hours
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Re: Culture storage [Re: x7x_x7x]
#8330241 - 04/26/08 11:37 AM (6 months, 23 days ago) |
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Quote:
x7x_x7x said:
http://www.shroomery.org/8509/Resurrecting-a-Better-Method-for-Long-Term-Storage-of-Mushroom-Cultures
"I came upon an article written by Michael D.Graham,a microbiologist at ATCC (1) that described the storage of yeast in sterile distilled water. What a brilliant idea! If that method stores yeast, It should work well on gourmet mushroom cultures, too. It's easy to do, very space efficient, allows the cultures to be stored at room temperature, and maintains thier viability for years. I contacted the author, he indicated that edible fungi stores even better than yeast, and you can store the spores as well as the hyphae often for decades!"
my fucking God what a data!
I already use this method for my yeast strains, I currently have over 30 stored in this manor and it works great! But Yeast has a dormant stage and once dormant do not require much, if any, food to survive. It seems that mycelium would need food or would eventually die.
Quote:
RogerRabbit said:
I put a hardwood stick in each slant, and they easily last 2 to 3 years under refrigeration. If the mycelium on the agar dies, it will still revive from the wood. I use medical tongue depressors.
If you don't use wood, make fresh slants every 6 to 8 months.
RR
This goes to reinforce my above statement, so I should add hard wood to the slant. Do I need to prepare the wood before adding it to the slant?
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Shop SMART, Shop S-MART!
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How to build a Martha
Self Healing Injection point lids
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How to build an Incubator
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x7x_x7x
urban farmer
Registered: 05/05/07
Posts: 679
Last seen: 1 hour, 6 minutes
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how about put some sterilized sawdust? will be OK, isn't?
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transhumanism + primitivism=magic mushrooms
SuctoSporeŽ
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lipa
Registered: 07/24/07
Posts: 585
Loc: Where it doesn't rain.
Last seen: 18 minutes, 58 seconds
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Re: Culture storage [Re: Cyber]
#8339804 - 04/28/08 10:21 PM (6 months, 20 days ago) |
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Cardboard soaked in myb works too!
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Ophanim
The Molecule'sSpirit
Registered: 10/01/07
Posts: 569
Last seen: 1 day, 15 hours
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Re: Culture storage [Re: lipa]
#8404653 - 05/15/08 09:49 AM (6 months, 4 days ago) |
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I don't know what a slant is, and I've never worked with agar, but can anyone tell me if a partially-colonized substrate such as Rye would work well for storage? Perhaps letting it colonize to 50%, then putting it in the fridge to purposely stall or slow its growth?
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lipa
Registered: 07/24/07
Posts: 585
Loc: Where it doesn't rain.
Last seen: 18 minutes, 58 seconds
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Re: Culture storage [Re: Ophanim]
#8404669 - 05/15/08 09:54 AM (6 months, 4 days ago) |
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What kind of culture are you wanting to store? Species
If you are going to use a grain to store your culture use millet from the health food store. use a small amount of it and let it colonize just to full capacity. Use a tube or small glass coke bottle stuffed with cotton. Cool it after that in your fridge. It will work fine. When you need it just take it out of the fridge, allow to sit for 24 hours then grab your loop and transfer some into what ever you want. return to fridge after that.
You don't have to use agar teks if you don't have access to the tools. You can also soak a piece of cardboard in some kind of non-simple sugar, sterilize in your bottle and inoculate. If you cut the cardboard into squares before you insert into your tube or bottle it will only require a set of tweezers to grab a square and inoculate something. It will last a good amount of time. "simple"
Edited by lipa (05/15/08 12:05 PM)
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x7x_x7x
urban farmer
Registered: 05/05/07
Posts: 679
Last seen: 1 hour, 6 minutes
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Re: Culture storage [Re: lipa]
#8404976 - 05/15/08 11:35 AM (6 months, 4 days ago) |
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invest few bucks and time in learning agar teks, agar is easy nd very useful tool (and toy)
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transhumanism + primitivism=magic mushrooms
SuctoSporeŽ
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shroober
Myco Junkie
Registered: 01/02/08
Posts: 782
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Re: Culture storage [Re: lipa]
#8405009 - 05/15/08 11:50 AM (6 months, 4 days ago) |
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I prefer using vacutainers. agar has a good write-up on it somewhere around here.
Basically just inject some LC into a vacutainer then let the myc settle at the bottom, then extract the nutrient LC out and replace with sterile water
The other benefit to this is vacutainers are super cheap if you get them from ebay. I just ordered 200 for $20 bucks
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MycoAu
Registered: 07/18/07
Posts: 647
Last seen: 1 hour, 53 minutes
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Re: Culture storage [Re: shroober]
#8405161 - 05/15/08 12:33 PM (6 months, 4 days ago) |
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Shroober, do you have any problems with cell death? Do draw up the LC-nutrient solution and replace with sterile water, only? or do you wash a few times with sterile water before allowing the last bit to remain in the vial. (Washing out all of the nutrients so that ONLY sterile water is left.)
Have you tried storing spores this way in place of a syringe?
Time limit for storage?
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shroober
Myco Junkie
Registered: 01/02/08
Posts: 782
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Re: Culture storage [Re: MycoAu]
#8405330 - 05/15/08 01:19 PM (6 months, 4 days ago) |
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Honestly I don't have long term experience with this method so I can't verify long term viability.
However, I believe agar said these would last a couple of years. I think RR's method might be best for LONG term storage, but for trade deals and 6-12 month storage I think vactuatiners make the most sense both in efficiency and expense.
Here's a thread where agar discusses using this method by getting myc water off of petri dishes...
http://www.shroomery.org/forums/showflat.php/Number/4094455#Post4094455
and another...
http://www.shroomery.org/forums/showflat.php/Number/3930928#3930928
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MycoAu
Registered: 07/18/07
Posts: 647
Last seen: 1 hour, 53 minutes
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Re: Culture storage [Re: shroober]
#8405487 - 05/15/08 02:06 PM (6 months, 4 days ago) |
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what volume of vacutainer are you using? I just might have to try this. (I'm tired of trying to keep up with agar storage methods.)
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shroober
Myco Junkie
Registered: 01/02/08
Posts: 782
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Re: Culture storage [Re: MycoAu]
#8405713 - 05/15/08 03:16 PM (6 months, 4 days ago) |
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10 ml
They also work great for storing spore solution
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Shroober's Lab | Rich Man's Pod | Martha | JumboTub | Outdoor Beds | Flow Hood | Bulk Substrate | Results
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fastfred
Registered: 05/17/04
Posts: 4,360
Loc: Dark side of the moon
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Re: Culture storage [Re: shroober]
#8420627 - 05/19/08 04:00 PM (5 months, 30 days ago) |
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Check out this thread:
http://www.shroomery.org/forums/showflat.php/Cat/0/Number/6886717
Fahtster has reported a successful storage method involving colonizing grains in jars with tyvek lids, which allows the grains to dry out. RR has also reported success reviving completely dried myc. Reviving dried myc from a sterile, mostly colonized grain jar should be a lot easier than from a specimen.
Otherwise slants and simply refrigerating LC works well as long as you revive and transfer them occasionally.
-FF
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_OttO_
PsychedelicTherapist
Registered: 06/01/05
Posts: 1,988
Loc: The fungal jungle
Last seen: 6 days, 14 hours
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Re: Culture storage [Re: fastfred]
#8421151 - 05/19/08 06:17 PM (5 months, 30 days ago) |
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Just chiming in to say I have stored a Liquid culture, in a cupboard (not under refrigeration), and used it successfully after just over a year. A PE6 liquid culture I had was still viable after 1.5 years, but it started to contaminate easily after this and lost lots of vigour.
And a spore print I used to get back into this hobby after going backpacking was left in an outdoor shed for 3 years, no special storage methods, and germinated without issue.
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Some travel photos - Noob friendly Liquid Culture - The story of the kangaroo poo cube
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420inmyapt
Stranger
Registered: 05/20/08
Posts: 9
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Re: Culture storage [Re: _OttO_]
#8432408 - 05/22/08 10:38 AM (5 months, 28 days ago) |
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Sweet, good info on storage, now i just need to get my hands on some prints... fucking CA laws 
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