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OfflineScavengerType
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Master Cultures without agar
    #9064452 - 10/11/08 07:24 PM (15 years, 5 months ago)

I've been reading up on senescence and the importance of keeping master cultures. However I do not have agar or petri dishes and didn't much want to get into agar at the moment. I was wondering what various methods people here would commonly use to keep a master culture without resorting to agar.


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"Have you ever seen what happens when a grenade goes off in a school? Do you really know what you’re doing when you order shock and awe? Are you prepared to kneel beside a dying soldier and tell him why he went to Iraq, or why he went to any war?"
"The things that are done in the name of the shareholder are, to me, as terrifying as the things that are done—dare I say it—in the name of God. Montesquieu said, "There have never been so many civil wars as in the Kingdom of God." And I begin to feel that’s true. The shareholder is the excuse for everything."
- Author and former M6/M5 agent John le Carré on Democracy Now.
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OfflineJuke Adro
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Re: Master Cultures without agar [Re: ScavengerType]
    #9064494 - 10/11/08 07:37 PM (15 years, 5 months ago)



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Someone said:  im actually not using ms, im using prints.
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OfflineScavengerType
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Re: Master Cultures without agar [Re: Juke Adro]
    #9064509 - 10/11/08 07:42 PM (15 years, 5 months ago)

I mean with regards to avoiding senescence.


--------------------
"Have you ever seen what happens when a grenade goes off in a school? Do you really know what you’re doing when you order shock and awe? Are you prepared to kneel beside a dying soldier and tell him why he went to Iraq, or why he went to any war?"
"The things that are done in the name of the shareholder are, to me, as terrifying as the things that are done—dare I say it—in the name of God. Montesquieu said, "There have never been so many civil wars as in the Kingdom of God." And I begin to feel that’s true. The shareholder is the excuse for everything."
- Author and former M6/M5 agent John le Carré on Democracy Now.
Conquer's Club

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Offlinenenl
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Re: Master Cultures without agar [Re: Juke Adro]
    #9064524 - 10/11/08 07:45 PM (15 years, 5 months ago)

I prefer that tek to MS LC, but there's not a way to isolate without agar, at least that I know of. A close substitute might be to noc the grain with a tissue of a good shroom to start with?


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hehe

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OfflineJuke Adro
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Re: Master Cultures without agar [Re: ScavengerType]
    #9064560 - 10/11/08 07:56 PM (15 years, 5 months ago)

Why do you want to keep a ms culture? when you can just get another print? if you want to keep isolates and store for a long time you need to learn agar and master culture slants with the tongue depressor, there is no other way about it IMO


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Someone said:  im actually not using ms, im using prints.
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Invisiblenoobieshroomie
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Re: Master Cultures without agar [Re: Juke Adro] * 1
    #9064575 - 10/11/08 08:03 PM (15 years, 5 months ago)

culture slants are the way to go

  -noobie-


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AMU

Best Thread Ever
CapZilla said:
not sure what GE and FAE are but i should probably get some.

Citric said:
Your signature is wrong on colonization temps!

GOOD JUDGMENT COMES FROM EXPERIENCE
EXPERIENCE COMES FROM BAD JUDGMENT

ROOM TEMP 70-75 IS BEST FOR COLONIZATION
Thank you mycochef

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OfflineScavengerType
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Re: Master Cultures without agar [Re: noobieshroomie]
    #9065021 - 10/11/08 10:00 PM (15 years, 5 months ago)

I was thinking of keeping grain jars for my master cultures and taking a spoon full at a time. I was wondering if anyone else ever did something like this.


--------------------
"Have you ever seen what happens when a grenade goes off in a school? Do you really know what you’re doing when you order shock and awe? Are you prepared to kneel beside a dying soldier and tell him why he went to Iraq, or why he went to any war?"
"The things that are done in the name of the shareholder are, to me, as terrifying as the things that are done—dare I say it—in the name of God. Montesquieu said, "There have never been so many civil wars as in the Kingdom of God." And I begin to feel that’s true. The shareholder is the excuse for everything."
- Author and former M6/M5 agent John le Carré on Democracy Now.
Conquer's Club

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Invisiblenoobieshroomie
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Re: Master Cultures without agar [Re: ScavengerType]
    #9065033 - 10/11/08 10:06 PM (15 years, 5 months ago)

how would you stop them from pining in side the jars?

    -noobie-


--------------------
AMU

Best Thread Ever
CapZilla said:
not sure what GE and FAE are but i should probably get some.

Citric said:
Your signature is wrong on colonization temps!

GOOD JUDGMENT COMES FROM EXPERIENCE
EXPERIENCE COMES FROM BAD JUDGMENT

ROOM TEMP 70-75 IS BEST FOR COLONIZATION
Thank you mycochef

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OfflineScavengerType
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Re: Master Cultures without agar [Re: noobieshroomie]
    #9065042 - 10/11/08 10:08 PM (15 years, 5 months ago)

the same way you stop petri dishes from fruiting, refrigeration.


--------------------
"Have you ever seen what happens when a grenade goes off in a school? Do you really know what you’re doing when you order shock and awe? Are you prepared to kneel beside a dying soldier and tell him why he went to Iraq, or why he went to any war?"
"The things that are done in the name of the shareholder are, to me, as terrifying as the things that are done—dare I say it—in the name of God. Montesquieu said, "There have never been so many civil wars as in the Kingdom of God." And I begin to feel that’s true. The shareholder is the excuse for everything."
- Author and former M6/M5 agent John le Carré on Democracy Now.
Conquer's Club

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OfflineP.Menace
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Re: Master Cultures without agar [Re: ScavengerType]
    #9065309 - 10/11/08 11:53 PM (15 years, 5 months ago)

Vacutainers and LC


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http://www.sporeworksgallery.com/Cubensae/Psilocybe_cubensis_Menace


roby000 said: thats true a shotgun is almost like a college degree in a sense that if you show it to the right person at the right time you could make a lot of money.

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Offlinesolumvita
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Re: Master Cultures without agar [Re: ScavengerType]
    #9065514 - 10/12/08 01:39 AM (15 years, 5 months ago)

You could in theory do that with the risk of contamination being reasonably high.
You would have to open the lid a lot more to get a spoonful out, which equals more chance of contam.  Whereas if you did the same thing with Agars tek and the LC your level of exposure is less as you can insert a needle into the bottle rather than a whole big dirty spoon.

Non-nutritive LC would be very similar to the use of distilled water and should give you a number of years of storage life.


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One of these days all the answers will be revealed until then we learn from each other!

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Invisibletripchip
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Re: Master Cultures without agar [Re: solumvita]
    #9065747 - 10/12/08 06:10 AM (15 years, 5 months ago)

hey how do you get the vaccuum out of the vacutainer? i know theres a tek around here about it.. since i had 60 cc's in a syringe i just let the vacutainer suck some out of the syringe. i got all these vacutainers and havent played with them yet. i know one day ill try to get it out and its gonna be a pain. i dont care if it goes to waste i did it for fun. mycelium water is a dime a dozen.


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Psilocybe Cubensis

Psilocybe comes from the Greek root "psilos" meaning bald head and Cubensis because it was first recognized as a new species from specimens collected in Cuba.

                                    :mushroom2:CHIP:mushroom2:

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Invisibletripchip
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Re: Master Cultures without agar [Re: ScavengerType]
    #9065752 - 10/12/08 06:16 AM (15 years, 5 months ago)

just do this tek here
then fill some test tubes up with the mycelium water. store in fridge.


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Psilocybe Cubensis

Psilocybe comes from the Greek root "psilos" meaning bald head and Cubensis because it was first recognized as a new species from specimens collected in Cuba.

                                    :mushroom2:CHIP:mushroom2:

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Offlineveda_sticks
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Re: Master Cultures without agar [Re: tripchip]
    #9065799 - 10/12/08 06:53 AM (15 years, 5 months ago)

Not really a master culture slant. The whole point of a master culture slant is storing a small peice of an isolated culture so that years down the line you can revve an isolated strain if you run out.

A test tube with LC in it, would be just a test tube of lc and not a master culture.

There are other teks that show how do agar using small jars and other ingredients.

Dont forget that a peice of wood in your masture culture slant will extend the life of your culture as the mycelium moves into the wood and lives there.


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PF TEK - writeup by EvilMushroom666
Lets Grow Mushrooms - RogerRabbit & RoadKills website with sample videos plus the full PF TEK video series. Alot of great information - BUY THE DVD
Cakes can and will pin! - So you think cakes suck for pins. Your wrong
Franks Simple Coir/Verm Tek
Franks Proper Pasturisation Tek
Franks Spawning To Bulk - Monotub
Professor Pinheads RTV Injection Port Tek
Foo Mans No Soak WBS Prep Tek

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OfflineP.Menace
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Re: Master Cultures without agar [Re: tripchip]
    #9065808 - 10/12/08 06:57 AM (15 years, 5 months ago)

Quote:

tripchip said:
just do this tek here
then fill some test tubes up with the mycelium water. store in fridge.




just let the vac-t suck the syringes down. they always fill up to a near perfect 10cc.
afterall they are hospital grade items


--------------------

http://www.sporeworksgallery.com/Cubensae/Psilocybe_cubensis_Menace


roby000 said: thats true a shotgun is almost like a college degree in a sense that if you show it to the right person at the right time you could make a lot of money.

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Invisiblesandman420
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Re: Master Cultures without agar [Re: veda_sticks]
    #9065966 - 10/12/08 08:42 AM (15 years, 5 months ago)

Quote:

veda_sticks said:
Not really a master culture slant. The whole point of a master culture slant is storing a small peice of an isolated culture so that years down the line you can revve an isolated strain if you run out.

A test tube with LC in it, would be just a test tube of lc and not a master culture.

There are other teks that show how do agar using small jars and other ingredients.

Dont forget that a peice of wood in your masture culture slant will extend the life of your culture as the mycelium moves into the wood and lives there.




It sure is a master culture. It is the same exact principle as a slant, just distilled water instead.

You then pull a few CCs out of the vaccutainer and noc up LCs with that.

One 9er tek or agar mycelium water will provide you with 50-70 vacutainers full of clone water, which could make thousands and thousands of 500ml LCs in days. The distilled water vacutainers will last in the fridge as long as a slant, or as long as you really want to.

If you use 9er tek its all clone, but if you do agars tek make sure the jar you suck out of has clone mycelium in it. Wallah no agar master cultures that last years.


--------------------
- Sandbag Tek - How To Sterilize Spawn Bags - All About Static Pressure / Pressure Drop for DIY Flow Hoods - Sandman's LC Tek-

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Re: Master Cultures without agar [Re: sandman420]
    #9066939 - 10/12/08 02:43 PM (15 years, 5 months ago)

if that's true I'll have to keep that in mind but my aim was to avoid senescence so the mycilium that comes from that method is all gen 1 mycilium thus making jars it inoculates gen 2. It is beginning to look like vacutainers are useful for long term culture storage, but I will have to store master cultures on agar. There doesn't seem to be another way.


--------------------
"Have you ever seen what happens when a grenade goes off in a school? Do you really know what you’re doing when you order shock and awe? Are you prepared to kneel beside a dying soldier and tell him why he went to Iraq, or why he went to any war?"
"The things that are done in the name of the shareholder are, to me, as terrifying as the things that are done—dare I say it—in the name of God. Montesquieu said, "There have never been so many civil wars as in the Kingdom of God." And I begin to feel that’s true. The shareholder is the excuse for everything."
- Author and former M6/M5 agent John le Carré on Democracy Now.
Conquer's Club

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Invisiblesandman420
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Re: Master Cultures without agar [Re: ScavengerType]
    #9067053 - 10/12/08 03:18 PM (15 years, 5 months ago)

Dude there is no way that you get senescence from expanding an original clone water onto an LC and then onto grain or BRF for spawn. It is still gen 1 going from LC to LC doesn't count as a generation. I don't know why you think you would get senescence? You store the culture in distilled water and blow out tons of LC for your inoculations.


--------------------
- Sandbag Tek - How To Sterilize Spawn Bags - All About Static Pressure / Pressure Drop for DIY Flow Hoods - Sandman's LC Tek-

Marijuanaut escapes earth to cultivate - Grow-room is church temple of the new stoner breed

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OfflineScavengerType
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Re: Master Cultures without agar [Re: sandman420]
    #9067430 - 10/12/08 05:29 PM (15 years, 5 months ago)

no but spawning in the jar creates 1 gen genetic degradation in the Myc water. If the method were able to somehow be used without using a significant amount of spawn growth, like in a small jar perhaps inoculated from the original LC and , I could see a working method on it that would produce a huge amount of LC that could be practically enough to call it a master culture. Additionally it would probibly take a few generations of this form of LC to degrade it enough to actually equal 1 normal generation, if I used your method of LC into LC I could preserve nominal senescence even longer. I can see how maybe this can make the whole practice insanely practical, but if I wanted to save strains to preserve their genetic integrity entirely I would have to do agar I think.

I am thinking I can do this easily without agar thanks for the advice.

Edited by ScavengerType (10/12/08 05:34 PM)

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Offlinecutemushie
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Re: Master Cultures without agar [Re: ScavengerType]
    #9170587 - 11/01/08 11:56 PM (15 years, 4 months ago)

Hi guys, would you please take a look at my long term culture storage method using grain and reanimation here:

http://www.shroomery.org/forums/showflat.php?Cat=0&Number=9170517&page=0&vc=1&PHPSESSID=#Post9170517 ;

I’m running into problems.  Would appreciate any comments.

Also what is meant by "hijacking a thread"?  I hope I haven't done it unintentionally here.  Sorry if I have.

Thanks.

Cheers,
cutemushie


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HAVE: Paneolus cyanescence, Paneolus cambodgiensis, Psilocybe cubensis, Flammulina velutipe (white, and brown Korean strain), Hericium erinaceous, Coprinus atramentarius, Coprinus comatus, Agrocybe aegerita, Pleurotus eryngii, Psilocybe subaeruginosa, Ganoderma lucidum, Pleurotus nebrodensis, Lepista nuda, Agaricus augustus, Pleurotus cystidiosus, Volvariella speciosa.

LOOKING FOR: Those that I don't have.

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