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OfflineTheAnimate
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Can someone help identify why I have contamination problems?
    #10133293 - 04/08/09 10:11 PM (14 years, 11 months ago)

I had 3 jars of PF mix and I was planning on using Eatyualive's Lazy Mofo Bag Tek( http://www.shroomery.org/forums/showflat.php/Number/1300760#1300760 ), then throwing it in a monotub with some coir.(to summarize) Anyway, I threw one jar in a bag with some uncolonized PF jars, and it got contaminated, with that forest green mold. Not super surprising. I mean, I did my best and wiped down my area with alcohol, and the bag as well, before throwing the jars in the bag, but when dealing in open air like that I really doubt there's anything you can do to ensure safety. But next time I planned to let the jar colonize longer, since I had thrown the colonized jar in the bag right as the jar looked to hit 100% colonization, before the mycelia had really taken hold of the substrate. So I thought that might help.

But I mixed up 2 more jars of PF mix anyway, for when my next jar was ready to be put into a bag. I put my PF mix into 2 pint jars, and put a lid on them, upside down, with no holes in it. After PCing these for an hour and a half at 10 PSI, I left the lids on and then put the jars into my incubator, because it seemed as good a place as any. Since the jars would stay closed, I figured there would be no harm in waiting around. But just a minute ago I checked my incubator, and those jars were infested with forest green mold. I'd already tried cleaning out my incubator from the last time I got contaminated, and I hadn't opened those jars at ALL. They were not 100% sealed or anything(the lid was upside down) and there was no dry vermiculite layer, but I really didn't expect to see them get contaminated.

Now, I'm not the most sterile person and as a result I've had to deal wiht a bit of contamination. But this one surprised me a lot, and now I'm going to try a lot harder. But I'm not even really sure what went wrong here. In the future, do you think I should seal these jars(keep the lid right side up) when PCing? Do you think I need to PC them for longer? I've already read the basics on keeping sterile, but do you guys think I should look into getting a flowhood, or something like that? I have already ordered some injection ports, and I'm going to look into both filter discs for jars and sheets of Tyvek, but if anyone has any more suggestions I'd be glad to hear them. I've already read the basics, I believe, but I promise I won't mind hearing them again, either. After all, I seem to have forgotten something.

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OfflineRogerRabbitM
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Re: Can someone help identify why I have contamination problems? (moved) [Re: TheAnimate]
    #10133315 - 04/08/09 10:17 PM (14 years, 11 months ago)

This thread was moved from Mushroom Cultivation.

Reason:
First of all, I wouldn't recommend a brf to brf transfer.  I have a super-clean lab with laminar flow hood, and I wouldn't even attempt it.

Second, if you're going to use the lids upside down, stick a piece of tyvek between the lid and the jar.  The metal to glass seal isn't perfect, and as the mix cools, it draws air into the jar, thus it draws any mold spores that are floating in the air right into your jars.  Tyvek should seal the gap.
RR

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OfflineTheAnimate
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Re: Can someone help identify why I have contamination problems? (moved) [Re: RogerRabbit]
    #10133849 - 04/09/09 12:22 AM (14 years, 11 months ago)

I will definitely try the Tyvek. I considered just using the lids normally, but they take time to unseal and I'd like to spend as little time as possible in the air. I realize there is no way to make this sterile, but I know others have had success with this method, and the possible payoff of fruiting a monotub compared to fruiting the two PF jars I have, I think is worth the risk. I believe the idea is that mycelium, having already colonized part of the substrate, has a better chance at spreading through the rest of the substrate before any mold you have introduced does. So while I know this is gonna be risky, I'd like to try it anyway, since I don't really have anything to lose. I'm just looking for tips on minimizing risk.

However I'm also looking at avoiding contamination in the future. I assume you're saying that BRF mix is more likely to get contaminated than other mixes? I've been meaning to look into others but I haven't gotten around to it. I'll probably try WBS next. It would make sense to me that BRF is more likely to get contaminated than other mixes because I've heard that BRF colonizes the fastest, and I guess that means that minimal exposures to contaminants can go much further, much faster, with BRF. I think I'll look into WBS tomorrow.

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