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zakzwijn
Stranger
Registered: 12/08/19
Posts: 55
Last seen: 1 year, 5 months
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Fully colonized healthy looking/smelling grain jar containing bacteria?
#27092012 - 12/16/20 11:38 AM (3 years, 1 month ago) |
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Hi fellow shroomers, so like many of you I was having this little unexpected issue with contamination. What I did:
- I prepared a sterilized master rye jar containing 4ml of spore syringe
- The rye jar fully colonized, smelled like mushrooms and looked healthy.
- I left the jar around for an extra week so the myc could took hold of the grain and eliminate possible contaminations (meh).
- I did g2g transfer from the colonized master jar to new sterilized rye jars. One teaspoon of grain and gave it a good shake.
- All the work was done under my 99.99% @0.3u laminar flow hood
- All jars were pc'ed for 90 minutes @15 psi and were fine.
- Some of the jars were a bit too wet, but not all of them.
So I was noticing slow growth... all my jars went bacterial :S I put what which I thought were bacteria under the microscope and bacteria were confirmed. All the jars didn't smell like mushroom as well, they all had that distinct sweet odor.
I found this thread with a comment from 'Machiavelliavore' stating:
Quote:
"An MS syringe inoculated jar is not a suitably clean source of inoculation for expansion. While the bacterial population may have been low enough in the master to produce decent growth, once you shake that bacteria all over a new jar, it can populate quite rapidly and prevent colonization, creating the impression of a stall.
I got fucked so hard by doing G2G's I thought were clean when I started. I suggest sticking to agar 2 grain or even straight MS 2 grain (along small grow scales) as a beginner, as it massively reduces the scale and time investment of your fuckups"
So I'm suspecting the master grain jar, because ALL my jars went bacterial. Is it a common occurrence what Machiavelliavore states? I expected my master grain jar to be clean, especially because it colonized very well... but that doesn't seem to be the case. I've seen other videos of people doing g2g transfers with much success, so maybe I'm just unlucky. What do you think?
Anyway, I'm going to do agar to grain transfer now.
Edited by zakzwijn (12/16/20 11:50 AM)
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HappinessStan
Fungivore



Registered: 10/10/12
Posts: 1,617
Loc: Worcester, UK
Last seen: 4 hours, 52 minutes
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Re: Fully colonized healthy looking/smelling grain jar containing bacteria? [Re: zakzwijn] 1
#27092067 - 12/16/20 12:07 PM (3 years, 1 month ago) |
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I think your last sentence answered your problems. Start with agar, ms syringes are almost always gonna be dirty. I just started using agar and it's so much easier than I thought it would be. I really struggled getting spores to germinate on agar before the bacteria did, which highlights just how unclean syringes are, this was from a reputable vendor. One thing I found really helped was making brf pucks in little glass jars, inoculating them through a self healing injection port (silicone, the cheap stuff works) and got mycelium in 7 days, then I just plucked some of that mycelium out in my SAB and put it to plates. The brf seems to speed up germination of spores whilst slowing down bacteria enough to start cleaning it up. Hope that helps. Good luck.
Edited by HappinessStan (12/16/20 12:09 PM)
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