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OfflinePowRGnome
No Stranger toCamus

Registered: 04/01/05
Posts: 83
Last seen: 12 years, 7 months
8 agar clones, no workspace, wwud?
    #8119573 - 03/08/08 11:56 AM (16 years, 10 months ago)

So I've got 8 clones on MEA in wide-mouth half-pints. I used some antibiotic to do them and they seem totally clean. They are (I guess what you'd call) first generation, as the original tissue is on the plates.

Can I transfer a wedge from these first generations into grain, or do I have to do another generation so the original tissue isn't on the plate? I would usually I guess do a two-from-one transfer onto more agar, but these were done in the bathroom and I don't want to push my luck any further without a glovebox or flowhood. Any help would be GREATLY GREATLY appreciated.

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Invisiblesmokedout420
The EnlightenedOne
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Registered: 12/16/07
Posts: 484
Re: 8 agar clones, no workspace, wwud? [Re: PowRGnome]
    #8119688 - 03/08/08 12:43 PM (16 years, 10 months ago)

you could but from what i been reading with agar,transfer,and isolation. you need to transfer until mycelium doesn't sector anymore. during that process you want to choose the ones with the most rhizo.

i've never done this process yet so i hope im telling you right i've been reading a lot on it lately, and gonna be trying soon enough.

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Offlinedizzy_jester
I aint noclown...

Registered: 03/16/07
Posts: 158
Last seen: 15 years, 10 months
Re: 8 agar clones, no workspace, wwud? [Re: smokedout420]
    #8119722 - 03/08/08 12:57 PM (16 years, 10 months ago)

Quote:

smokedout420 said:
you could but from what i been reading with agar,transfer,and isolation. you need to transfer until mycelium doesn't sector anymore. during that process you want to choose the ones with the most rhizo.

i've never done this process yet so i hope im telling you right i've been reading a lot on it lately, and gonna be trying soon enough.




when working with agar you want to isolate a single sub strain...usually the fastest growing and the most rhizmorphic... usually takes a few times until you reach there going from agar to agar.

just keep cutting out the big parts and transfering until it just spreads into a circle and uniform

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OfflinePowRGnome
No Stranger toCamus

Registered: 04/01/05
Posts: 83
Last seen: 12 years, 7 months
Re: 8 agar clones, no workspace, wwud? [Re: dizzy_jester]
    #8119750 - 03/08/08 01:05 PM (16 years, 10 months ago)

It is an isolate since each is from the stem of a fruitbody. It's growing very rhizo and very quickly, it's actually about to start climbing the walls of the jars. Here's what I think are my options:

Option 1:

Transfer a wedge from each plate to 8 different pint jars of grain, incubate.


Option 2:

Transfer a wedge from each plate to 8/16 different half-pints of MEA, incubate, then transfer from those to pint jars of grain.

I'm leaning towards option 1 because I think with semi-sterile bathroom conditions the grain is likely to be more selective. However, the thing I'm worried about is using the plates that have the original stem tissue on them. Is this an issue?

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Invisiblesmokedout420
The EnlightenedOne
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Registered: 12/16/07
Posts: 484
Re: 8 agar clones, no workspace, wwud? [Re: PowRGnome]
    #8119987 - 03/08/08 02:18 PM (16 years, 10 months ago)

options you can got. this way you can save your agar.

agar 2 lc
LC Cloning - Newbie friendly pictorial tek!

or
faht's iso/ myc syringe tek
homemade lids that have stood the test of time

or

agar 2 grain



if you dont have a flowhood at least use a glove box. or take ur chances and shut the heat off for a hour. let the dust settle in ur home got to the bathroom take everything ur gonna use and clean the shit out of the bathroom. then work in there. i have a buddy that uses that method all the time and tells me to quit saving for a flowhood but id rather get the flow hood.

Edited by smokedout420 (03/08/08 03:31 PM)

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