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OICU812
NC Tree Farm owner


Registered: 11/06/11
Posts: 1,064
Loc: Foothills of NC
Last seen: 3 years, 4 months
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Stabs
#18875117 - 09/22/13 09:07 AM (10 years, 4 months ago) |
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In perusing Dr. Raines-Casselman's article "Yeast Propagation and Maintenance: Principles and Practices" for the upteenth time, I payed particular attention to the last section entitled: "Methods of Yeast Maintenance" and for seemingly the first time noticed the paragraph entitled Stabs. It occurred to me that creating small centrifuge tubes (1.5 -to- 10 ml) with a media as described in the cited paragraph and injecting a small amount of mycelium from a syringe for long term storage might have merit. It also occurred to me that if I could repeatedly successfully cover the mycelium in slants with a slurry of media as described, it might result in extending slant storage time.
Excerpted from Dr. Raines-Casselman's article "Yeast Propagation and Maintenance: Principles and Practices": "Stabs- Stabs are upright tubes or bottles of semi-solid agar media. They contain only 0.7-1.0% agar. These are best prepared in screw-cap container with rubber inserts in the caps such as bijou, McCartney or Universal bottles but standard culture tubes may also be used. Yeast is applied with an inoculation loop and is inserted or stabbed into the agar all the way to the bottom of the tube or bottle. The stab is incubated and stored similar to a slant. Stabs are a common method used for long term storage of bacteria. Like plates, I have not seen any reference to this technique for yeast. The main advantage of this procedure is that it minimizes exposure to air which appears to be the primary limiting factor for shelf-life and stability. The disadvantage is that they are difficult to work with since the yeast is embedded in the agar and difficult to see. Also a fair amount of agar is usually picked up during transfer. It is unclear what the shelf-life is for stabs but my guess is that it would be at least 2-4 years (using tight-sealing bottles). Stabs therefore are a good for long-term storage, but not appropriate for routine propagation."
Further reading on Agar and Culture Storage
-------------------- -------------- "Democracy is two wolves and a lamb voting on what to have for lunch. Liberty is a well-armed lamb contesting the vote!" --Benjamin Franklin "Those who give up liberty for security won't have, or deserve, either.". . . Benjamin Franklin ----> Read: The Fight of our Lives - Defeating the Ideological War Against the West - by Victor Davis Hanson
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hidyn
Noob



Registered: 07/14/13
Posts: 268
Loc: Canada
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So this is similar in principle to long term storage in dH20 because tissue is suspended in an environment completely devoid of nutrients and air exchange, yes?
...I need to start culturing my own yeast.
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thelorax121
Stranger

Registered: 10/12/05
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Re: Stabs [Re: hidyn]
#18878126 - 09/22/13 09:45 PM (10 years, 4 months ago) |
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Because he describes an incubation time and storage procedure similar to slants, I am assuming the "semi-solid agar media" is enriched with something (MYEA, PDYA, CMYA etc) allowed to colonize, then put into storage. I would appreciate some other folks to chime in though, as this does seem like it would be a bit easier to prepare than slants, given less media is used.
-------------------- Greens for all, and to all a good greens!
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OICU812
NC Tree Farm owner


Registered: 11/06/11
Posts: 1,064
Loc: Foothills of NC
Last seen: 3 years, 4 months
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Table 2 in the linked text above describes two formulas used for slants, plates and limited liquid propagation.
-------------------- -------------- "Democracy is two wolves and a lamb voting on what to have for lunch. Liberty is a well-armed lamb contesting the vote!" --Benjamin Franklin "Those who give up liberty for security won't have, or deserve, either.". . . Benjamin Franklin ----> Read: The Fight of our Lives - Defeating the Ideological War Against the West - by Victor Davis Hanson
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forrest



Registered: 11/16/12
Posts: 1,011
Loc: The Netherlands
Last seen: 4 years, 6 months
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very interesting site! and the part about the stabs.
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Edited by forrest (09/24/13 11:30 AM)
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fastfred
Old Hand



Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
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Yea for stabs. Not advanced though, this is micro 101 crap.
-------------------- It drinks the alcohol and abstains from the weed or else it gets the hose again. -Chemy The difference between the substances doesn't matter. This is a war on consciousness, on our right to the very essence of what we are. With no control over that, we have no need to speak of freedom or a free society. -fireseed "If we are going to have a war on marijuana, the least we can do is pull the sick and the dying off the battlefield." -Neal Levine (MPP) I find the whole "my drug should be legal but yours should be illegal" mindset disgusting and hypocritical. It's what George Bush and company do when they drink a cocktail and debate the best way to imprison marijuana users. -Diploid
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Amanita virosa
botanist by day


Registered: 12/04/11
Posts: 2,458
Loc: north kakalacky
Last seen: 6 months, 17 days
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yeast, being unicellular, would lend themselves to this type of storage better than filamentous fungi, it would seem. Sterilized vegetable oil works great, poured over the top of a slant for long term storage. I woke up some five year old cultures of morel that were stored this way. the trick is having a good fridge that keeps them just above freezing, but with out freezing them. as we all know micro fridges suck at this. I am fortunate to have a 4000$ fancy culture storage fridge at work to keep my long term storage in. otherwise, every six months I dip in, grow out, reslant.
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OICU812
NC Tree Farm owner


Registered: 11/06/11
Posts: 1,064
Loc: Foothills of NC
Last seen: 3 years, 4 months
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Quote:
Amanita virosa said: yeast, being unicellular, would lend themselves to this type of storage better than filamentous fungi, it would seem. Sterilized vegetable oil works great, poured over the top of a slant for long term storage.
Very interesting! I'm always up for trying something new once, twice if I like it!
So when you revive the culture, do you drain out the oil or just retrieve the mycelium through it?
-------------------- -------------- "Democracy is two wolves and a lamb voting on what to have for lunch. Liberty is a well-armed lamb contesting the vote!" --Benjamin Franklin "Those who give up liberty for security won't have, or deserve, either.". . . Benjamin Franklin ----> Read: The Fight of our Lives - Defeating the Ideological War Against the West - by Victor Davis Hanson
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Amanita virosa
botanist by day


Registered: 12/04/11
Posts: 2,458
Loc: north kakalacky
Last seen: 6 months, 17 days
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Dump the oil. Then sub out. It takes the Myc a while to revive. Morel grows fast but it s till took maybe a week before it broke loose of the residual oil and grew. I have not tested my other slants for viability but I do have some that are going on five years. Figure I will leave them till I need them.
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Amanita virosa
botanist by day


Registered: 12/04/11
Posts: 2,458
Loc: north kakalacky
Last seen: 6 months, 17 days
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