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OfflineTerry M
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LC for short-term culture in half pint jars * 1
    #14202324 - 03/29/11 05:06 AM (12 years, 11 months ago)

There's method to my madness. I've learned the hard way that LC is not a good long-term storage medium. Most of my old refrigerated jars are now dead. Plus and more importantly, contaminants may not visible in LC.

So my master plan is to use LC on a short-term basis:

1. Maintain a set of refrigerated agar slant cultures, which are good up to 2 years. I use the wooden stick method on RR's DVD.

2. Inoculate a few plates with a bit of the slant culture, let them grow out, and refrigerate.

3. When you want to grow some particular shrooms for a while, take a plate out of the fridge and let it sit at room temperature to reactivate the mycelium.

4. PC a half pint standard mouth jar filled with LC, as well as a blender base. The role of the latter will become obvious.

5. Drop a wedge of agar into LC jar, and blend it in a standard Oster blender.* This breaks up the agar mycelium into tiny pieces (which cannot be done by shaking with a marble or stirring -- see page 139 of Stamets' GGMM).

6. Put back the standard LC lid on the jar using a flow hood or glove box. You now have a completely fresh and alive, and (very very probably) contaminant-free LC to inoculate MANY grain jars with for the next 3 months or so! And a half pint is equivalent to 18 10ml syringes, plus stores conveniently in the frig. When you want to repeat the cycle, or get another mushroom strain going, go to step 3.


One of my half pint standard mouth LC jars.

* Blenders are pretty much all designed to be compatible with a standard mouth canning jar. This is not mere coincidence, but a decades-old design to aid the "homemaker" to blend up the contents of canning jars. The Oster, however, has all-metal base and blades, so it can be sterilized in the PC without melting.


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Liberté, égalité, humidité.

Edited by Terry M (03/29/11 10:13 AM)

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OfflineSWIM Jr
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Registered: 02/14/11
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Re: LC for short-term culture in half pint jars [Re: Terry M]
    #14202382 - 03/29/11 05:45 AM (12 years, 11 months ago)

Very Nice.

Have you tried doing LC to LC every few months? I have some going for 6 months now, with MEA and honey.

Do you use plain water to make the LC with the agar plate?


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OfflineTerry M
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Re: LC for short-term culture in half pint jars [Re: SWIM Jr]
    #14202447 - 03/29/11 06:23 AM (12 years, 11 months ago)

Quote:

SWIM Jr said:
Very Nice.

Have you tried doing LC to LC every few months? I have some going for 6 months now, with MEA and honey.

Do you use plain water to make the LC with the agar plate?




Thanks, SWIM. I haven't actually done this yet -- just assembling what I need. I know I can keep LC going for 6 months, but I want to be safe and do slant/plate backups for longer term. I waited almost a year before going back to my quart LC jars, and had an unpleasant surprise.

And I'm going to use tap water for the LC medium. I'm following GGMM pages 137-141, but using just a single LC step instead of the 2-phase one in in the book. Stamets procedure is for high-volume commercial growers, and uses a metal Eberbach to bust up the agar in sterilized water, and then a transfer to some malt sugar LC Erlenmeyers for stirring and growth.

It's gonna be hard to say goodbye to my quart LC jars in the fridge, but they're giving way to a more compact and I think more reliable (especially for forgetful people like myself) solution.


--------------------
Liberté, égalité, humidité.

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OfflineTerry M
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Re: LC for short-term culture in half pint jars [Re: Terry M]
    #14202680 - 03/29/11 08:24 AM (12 years, 11 months ago)

I'm going to do this within a week or two, and post a detailed tek with lotsa picture! This is really just a variation/elaboration of the old 9er Tek.

Note that this tek is designed for people who are already maintaining their cultures on agar, but instead of having to make a grain master and do grain-to-grain transfers which use up your master at once, have:

1. An LC "master" which is made directly from agar, and replaces the grain master. Like a grain master, it is just one generation removed from agar.

2. A fresh master which lasts months, and can inoculate around a hundred grain jars.

3. The convenience and ease of LC grain jar inoculation.


--------------------
Liberté, égalité, humidité.

Edited by Terry M (03/29/11 08:57 AM)

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InvisibleDoc_T
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Re: LC for short-term culture in half pint jars [Re: Terry M]
    #14204893 - 03/29/11 05:25 PM (12 years, 11 months ago)

If I can suggest another method that gets you to the same result with less work and less risk... agar->grain->grain lc.


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OfflineTerry M
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Re: LC for short-term culture in half pint jars [Re: Doc_T]
    #14205023 - 03/29/11 05:53 PM (12 years, 11 months ago)

Quote:

Doc_T said:
If I can suggest another method that gets you to the same result with less work and less risk... agar->grain->grain lc.



Wow, I just read up on GLC, which I knew nothing about (or if I did, forgot it). It's just great! Thanks, Doc T!

As I understand it, the GLC will keep in the fridge on the order of months, just like a sugar-nutrient LC. So I could use my little LC jars for storing this in the fridge, and when I want a syringe to noc some grain, I just use the self healing injection port. And the Whatman filter allows all the gas exchange needed in the frig, right?


--------------------
Liberté, égalité, humidité.

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Re: LC for short-term culture in half pint jars [Re: Terry M]
    #14205145 - 03/29/11 06:18 PM (12 years, 11 months ago)

I store it in sealed syringes in the fridge in ziploc bags.
Then bring it out, shoot a tiny bit into grain masters (I use pints) and then do g2g when it colonizes.
So it lasts a much longer time and you check it for purity at each use.


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You make it all possible. Doesn't it feel good?

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OfflineTerry M
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Re: LC for short-term culture in half pint jars [Re: Doc_T]
    #14205207 - 03/29/11 06:32 PM (12 years, 11 months ago)

So you store several GLC syringes in the fridge for a while? I'm trying to avoid the large number of syringes I'd need to store my soon-to-be-20 different edible varieties. Yes, I know I am a nut to maintain this many cultures. :grin:

But I could use a variation of your method to store GLC in my half pint jars? When when I want to use one, I suck out some liquid with a syringe, noc a small grain master, and do a G2G transfer just like you are doing.

Would this work, or am I misunderstanding something?


--------------------
Liberté, égalité, humidité.

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Re: LC for short-term culture in half pint jars [Re: Terry M]
    #14205244 - 03/29/11 06:38 PM (12 years, 11 months ago)

Syringes are smaller than jars. But I do it this way to keep each culture clean.
If you keep re-tapping the same jar, you risk damaging your entire lineage.
But if you split it up at harvest and store them, you have backup.


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You make it all possible. Doesn't it feel good?

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OfflineTerry M
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Re: LC for short-term culture in half pint jars [Re: Doc_T]
    #14205278 - 03/29/11 06:45 PM (12 years, 11 months ago)

Quote:

Doc_T said:
Syringes are smaller than jars. But I do it this way to keep each culture clean.
If you keep re-tapping the same jar, you risk damaging your entire lineage.
But if you split it up at harvest and store them, you have backup.



This sounds like the right way to go. I could simply store a few syringes of one variety in the same plastic bag in the fridge. All together they wouldn't take up that much space.

Well, making the LC jars was fun. But I think I'll be ordering a lot of syringes soon! Thanks for the great input, Doc T!


--------------------
Liberté, égalité, humidité.

Edited by Terry M (03/29/11 06:46 PM)

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Re: LC for short-term culture in half pint jars [Re: Terry M]
    #14205360 - 03/29/11 07:00 PM (12 years, 11 months ago)

I like 30 mL syringes for liquid masters. They fit nicely in the carrot drawer of your fridge. :thumbup:


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OfflineOldYamaha
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Re: LC for short-term culture in half pint jars [Re: Doc_T]
    #22601400 - 12/01/15 08:58 PM (8 years, 2 months ago)

Quote:

Doc_T said:
If I can suggest another method that gets you to the same result with less work and less risk... agar->grain->grain lc.




Just adding this to my favorites. No comments.


--------------------
Semi Corporate World :smirk:Retired and looking for a new part time hobby business, just to stay off my wife's bad side.:thumbup::thumbup:
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InvisibleLocN9ne
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Re: LC for short-term culture in half pint jars [Re: OldYamaha]
    #22601424 - 12/01/15 09:02 PM (8 years, 2 months ago)

Quote:

OldYamaha said:
Quote:

Doc_T said:
If I can suggest another method that gets you to the same result with less work and less risk... agar->grain->grain lc.




Just adding this to my favorites. No comments.



You should try making a bookmark, instead of flossing your resurrection skills.


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OfflineOldYamaha
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Re: LC for short-term culture in half pint jars [Re: LocN9ne]
    #22603521 - 12/02/15 10:42 AM (8 years, 2 months ago)

Quote:

LocN9ne said:
Quote:

OldYamaha said:
Quote:

Doc_T said:
If I can suggest another method that gets you to the same result with less work and less risk... agar->grain->grain lc.




Just adding this to my favorites. No comments.



You should try making a bookmark, instead of flossing your resurrection skills.




"You should try making a bookmark?" How???:takingnotes:
"instead of flossing your resurrection skills." What???:confused:


--------------------
Semi Corporate World :smirk:Retired and looking for a new part time hobby business, just to stay off my wife's bad side.:thumbup::thumbup:
Favorite Bumpersticker!
Smile, Jesus Loves You!:cool:
Everyone else thinks your an asshole

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