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madis503
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Registered: 07/29/09
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MEYA problem
#16339789 - 06/06/12 05:52 AM (12 years, 7 months ago) |
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Hi all! i tried cloning with cardboard and it didnt work out, so now i did it with agar and it seems that i have failed again. the mushroom tissue has not grown. on some plates i see bacteria growing but no mycelium on any of the dishes. the recipe was 200ml water, 4 g malt, 3,5g agar agar, 1g yeast. i suspect that i bought something malt-like? Does it has to be powder too?

 yeast and agar:

i dont have parafilm yet but i ordered a roll and a box of 3 sector petri dishes (480) i just dont want to fail anymore. those dishes have been sitting for 4 days now  these are sealed with micropore tape.
 

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Psy&Goa
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: MEYA problem [Re: madis503]
#16339821 - 06/06/12 06:22 AM (12 years, 7 months ago) |
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You are successfully growing bacteria, that's a good first step. What kind of tissue cloning are you doing? A wild specimen?
-------------------- You make it all possible. Doesn't it feel good?
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madis503
psühhonaut


Registered: 07/29/09
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Re: MEYA problem [Re: Doc_T]
#16339982 - 06/06/12 07:32 AM (12 years, 7 months ago) |
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no no, just picked 2 of the ones i liked the most and cloned them, but they were in the refridgerator for 4 days. but do i have the right malt?  and if theres bacteria growing then myc wont grow at all? and can i re melt the agar what was left over?
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Psy&Goa
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: MEYA problem [Re: madis503]
#16339998 - 06/06/12 07:38 AM (12 years, 7 months ago) |
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Quote:
madis503 said: but do i have the right malt? 
You have maltose, refined malt sugar. It will work ok, especially with the added yeast.
Quote:
madis503 said: and if theres bacteria growing then myc wont grow at all?
Maybe myc will grow and if so you can transfer it to a clean dish. But the fact that bacteria are growing means the agar works.
Quote:
madis503 said: and can i re melt the agar what was left over?
If it's clean and not growing anything, yes.
-------------------- You make it all possible. Doesn't it feel good?
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madis503
psühhonaut


Registered: 07/29/09
Posts: 466
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Last seen: 2 years, 30 days
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Re: MEYA problem [Re: Doc_T]
#16340272 - 06/06/12 09:13 AM (12 years, 7 months ago) |
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thank you
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Psy&Goa
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madis503
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Re: MEYA problem [Re: madis503]
#24256659 - 04/19/17 09:23 AM (7 years, 8 months ago) |
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that recipe has worked for a long time, but about a year i'm having growth problems on this. is it possbible that the maltose is old? i tried potato flake agar also, didn't grow anything, even bacteria were slow.. maybe it's the additives in the potato flakes? also maybe the yeast moisture additive (E491) is bad? Damn i dont know what to look at anymore..
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Psy&Goa
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,915
Loc: Milky way
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Re: MEYA problem [Re: madis503]
#24256679 - 04/19/17 09:29 AM (7 years, 8 months ago) |
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That stuff doesn't really go bad. It just absorbs moisture and gets clumps
Judging by the last four pictures in the first post your problems are coming from being not so good with agar
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madis503
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yeah that was 4 years ago... i've tried with powder maltodextrin also, but it's the wrong stuff i guess, is maltodextrose ok? i think i can get it now from a shop... i'm just boiling a new bottle of the original formula I used, what used to grow nice myc. Hoping it will work... really want to make some slants.
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Psy&Goa
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
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Re: MEYA problem [Re: madis503]
#24256722 - 04/19/17 09:45 AM (7 years, 8 months ago) |
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Maltodextrine is maltodextrose
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madis503
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strange, it didn't work good. Is it possible to overcook the mixture somehow?
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Psy&Goa
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,915
Loc: Milky way
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Re: MEYA problem [Re: madis503]
#24257083 - 04/19/17 11:41 AM (7 years, 8 months ago) |
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Yes, don't go much more than 25-30m at 15psi. It only takes 15m to sterilize loads under 1L in volume for most of the basic agar media
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madis503
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Quote:
bodhisatta said: Yes, don't go much more than 25-30m at 15psi. It only takes 15m to sterilize loads under 1L in volume for most of the basic agar media
yeah that might be the problem, i watched LGM videos and they suggested to pc for 1.5 hours i think, if i remember right. so the last time it cooked long... but that doesn't explain the failures before, i trasnfered wedges from older dish and it just wouldn't grow Anyway I'll comment when i see growth on my new dishes.
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Psy&Goa
Edited by madis503 (04/19/17 12:43 PM)
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,915
Loc: Milky way
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Re: MEYA problem [Re: madis503]
#24257498 - 04/19/17 02:35 PM (7 years, 8 months ago) |
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1.5 hours is for grain jars. but I do 2 hours
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madis503
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Quote:
bodhisatta said: 1.5 hours is for grain jars. but I do 2 hours
yes i know that also, but thank you for answering. Anyway, i guess the problem is i overcooked the mix.. But the new batch is good, I put some colonized grain on it, grows well and already made some transfers.
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Psy&Goa
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