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Myc097
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Registered: 11/11/18
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Please help!
#26836545 - 07/21/20 04:41 PM (3 years, 6 months ago) |
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Hey guys so I have been having issues with agar and I am just about over it. I have tried a2g countless times with clean agar similar to the picture below to just get bacterial spawn. So in a test I prepped 20 grain jars ( 45minute soak in boiling water-pc for 90 minutes) and let them sit the only issue that I found over a month later was 1/20 developing trich. So it’s not my spawn or my inoculate what would be causing the issue. I feel like if I’m getting clean plates it’s also hard to say it’s sterile tek? Please help I’m literally going insane over here. Thank you for any input it’s much appreciated!
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bodhisatta 
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Re: Please help! [Re: Myc097]
#26836563 - 07/21/20 04:50 PM (3 years, 6 months ago) |
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Spawn looks bad. Plate looks good
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skullhuman
the skullman cometh



Registered: 06/15/09
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Soak the grains overnight to germinate bacterial endospores, so that you can get rid of them when you boil/PC. If that's not where the problem is being introduced, then it's your technique when going from agar to grain.
Edited by skullhuman (07/21/20 05:55 PM)
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Myc097
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Could you point me to a description on the best sterile tek that has to be it.
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skullhuman
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Re: Please help! [Re: Myc097]
#26836797 - 07/21/20 06:36 PM (3 years, 6 months ago) |
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The best rye tek? No need to reinvent the wheel, just follow this classic and your jars will always be prepped perfectly:
What is your technique when going from agar to grain?
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bodhisatta 
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Quote:
skullhuman said: Soak the grains overnight to germinate bacterial endospores, so that you can get rid of them when you boil/PC. If that's not where the problem is being introduced, then it's your technique when going from agar to grain.
I haven't soaked grains in ages.
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skullhuman
the skullman cometh



Registered: 06/15/09
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That may not be sufficient for op. Although I suspect his transfer technique is probably what's off.
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Myc097
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Spray down SAB with soapy water spray jars and gloves down with 70%, loosen jar lid, cut agar, place in spawn jar. Never putting hand over agar or spawn jar.
Where do you even find rye? I’ve been using bodhis oat tek since I started.
Edited by Myc097 (07/21/20 10:31 PM)
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KuroNeko
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Registered: 05/19/20
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Endospore germination has been debunked. No need to soak but I do soak because I don't like burst grains as they stick to sides of a jar and ruin aesthetics when shaking.
Read here: https://www.shroomery.org/forums/showflat.php/Number/21604725
-------------------- English not my native language, please excuse grammar.
 
Edited by KuroNeko (07/21/20 11:27 PM)
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A.k.a
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Re: Please help! [Re: KuroNeko]
#26837272 - 07/21/20 10:59 PM (3 years, 6 months ago) |
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I had trouble with knocking up jars at first too. Same deal, clean plates but botched the inoculation.
Try to move slowly, open the lid before so it’s loose and easy to lift up, and try to keep the wedge barely dangling from the scalpel so you can just crack the lid and drop it in real quick. Fiddling around getting the wedge off is what got me a lot of times.
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LAGM2020     
Edited by A.k.a (07/21/20 10:59 PM)
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KuroNeko
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Re: Please help! [Re: A.k.a]
#26837319 - 07/21/20 11:36 PM (3 years, 6 months ago) |
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Quote:
A.k.a said: Try to move slowly, open the lid before so it’s loose and easy to lift up, and try to keep the wedge barely dangling from the scalpel so you can just crack the lid and drop it in real quick. Fiddling around getting the wedge off is what got me a lot of times.
Yes, don't touch receiving jar rim with scalpel, don't touch agar wedge with the rim, don't hover your hand over open plate and jar, don't keep plate or jar open for unnecessary amount of time. Like he said: lift agar piece with tip of your scalpel, move it near receiving jar, crack the lid open, drop agar and close immediately. Scalpel should only be as far above as length of blade. I lift my agar wedges in a way I need to only rotate my scalpel to drop the wedge. You can practice it with a dirty agar plate. Stab wedge with your blade horizontally, lift and when you rotate blade to be vertically the wedge should just slide off the blade easily. I use #11 blades with #3 handle like most people. Other curved blades may not be able to do this.
-------------------- English not my native language, please excuse grammar.
 
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bodhisatta 
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Registered: 04/30/13
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Quote:
skullhuman said: That may not be sufficient for op. Although I suspect his transfer technique is probably what's off.
Its sufficient for anyone with a pressure cooker. You know the thing that kills endospores.
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Myc097
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Re: Please help! [Re: KuroNeko]
#26837539 - 07/22/20 05:00 AM (3 years, 6 months ago) |
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Thank you for the detail truly! That must be the issue I’m hitting the side of the jar to knock off the wedge every time.
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bodhisatta 
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Registered: 04/30/13
Posts: 61,890
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Re: Please help! [Re: Myc097]
#26837541 - 07/22/20 05:04 AM (3 years, 6 months ago) |
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I use the lip of the jar to get the wedge off the scalpel.
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AtmozFear
just a shade of myself


Registered: 01/25/19
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Quote:
skullhuman said: Soak the grains overnight to germinate bacterial endospores, so that you can get rid of them when you boil/PC. If that's not where the problem is being introduced, then it's your technique when going from agar to grain.
No one ever mentions this. I saw this also in some grain-soak tek (can't remember which one) and I think this is great advice, tho I don't think it is this particular OP's problem.
Edited by AtmozFear (07/22/20 05:06 AM)
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Myc097
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Am I just really shitty at this? Hahaha I started doing ms to lc and wanted to do it right and now I have just straight failures. I try to follow bods videos but maybe I’m just shit at it lol
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AtmozFear
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Re: Please help! [Re: Myc097]
#26837561 - 07/22/20 05:45 AM (3 years, 6 months ago) |
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practice makes perfect
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A.k.a
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The endospore thing is another one of the many things debated around here. From the stuff I’ve read I’m convinced it’s not a factor and it’s just old info hanging around, but that’s just me.
I sometimes use the lip to knock the wedge off too but I think there’s a way to do that. Still keep as little of the scalpel over the mouth as possible and pull it off in a smooth motion. I used to do it where half the handle would be over or in the jar, now I make sure to just have a tiny part of the blade over the opening while lining up the lip with the wedge to bump it off.
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LAGM2020     
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skullhuman
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Re: Please help! [Re: A.k.a]
#26838138 - 07/22/20 11:57 AM (3 years, 6 months ago) |
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Quote:
bodhisatta said:
Quote:
skullhuman said: That may not be sufficient for op. Although I suspect his transfer technique is probably what's off.
Its sufficient for anyone with a pressure cooker. You know the thing that kills endospores.
I've been doing things since 09, when the prevailing logic was that hard endospores designed to weather harsh conditions can survive pressure cooking if not first germinated. I stopped reading grain teks a long time ago because that is how long I haven't needed them for, so please forgive me if I missed a memo or two sir.
Quote:
A.k.a said: The endospore thing is another one of the many things debated around here. From the stuff I’ve read I’m convinced it’s not a factor and it’s just old info hanging around, but that’s just me.
Maybe. Or maybe RR was right about the soak thing and whenever a noob has endospore problems a bunch of guys rush in to tell him that that theory has been 'debunked' and to not bother. Or maybe not, and it's just his agar technique that's off. I haven't personally examined soaked vs unsoaked endospores that have been thru PC cycles under a microscope so I can't be certain in either case.
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