beluga said:

So, spore syringes are inherently dirty. But that doesn't mean that direct inoculation can't work, but it's a much bigger uncertainty. I did direct inoculation for a while with some success but also a lot of green.

Couple questions, (I think you may have posted, but I suck at going back and forth on my phone):
- Did you vent your PC for 10 minutes once it had a full stream of steam?
- How long did you PC?

Your grain looks prepped quite well, but that doesn't make or prevent contamination... it had to be introduced somewhere.

Couple notes:
- Glove boxes can create a positive air pressure within them that will force stuff into your media. I would pop those gloves out and try it in the Still Air Box form. Deliberate, smooth movements so to not disturb the air much.
- Did I see that you had tape over your grain jar lids? They have to be able to release steam in order to properly sterilize, so there should be some form of air exchange.

Any extra details you can think of, I'm happy to assist in troubleshooting.

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beluga said:

So, in your lids, you wanna have a ~1/4" hole that has a filtering material - like polyfill, tyvek, micropore tape, or synthetic filter discs made from PTFE - over it. This prevents microbial transfer and allows gas exchange.

The process:
- load jars with prepped grain
- put lids on and loosen them a little bit
- cover in foil to prevent water from dripping through which could throw off your grain moisture
- fill PC with 3 quarts of water (per Presto 23 quart) and bring to a boil
- put your lid on without the rocker and wait until there is a steady stream of steam coming out of the valve.
- vent for 10 minutes.
- put your rocker on and bring to 15 PSI
- PC for 90 minutes to 120 minutes at 15 PSI.
(Pro tip: you can attach 2 US quarters to your rocket to bring it to 17-18PSI without releasing steam... this will make 90 minutes more effective.)
- let it come to atmospheric pressure on its own
- open, tighten the jar lids, and remove foil.

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beluga said:

Gypsum..?
Looks a little heavy.. but I've only tried using it a few times - usually in soak/boil water.

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beluga said:

Micropore tape under magnification:

40x
View attachment 1056250

100x:
View attachment 1056251

400x:
View attachment 1056252

There's also the comparison of a P. cubensis spore in relation to those gaps somewhere that I'm having trouble finding.
The gaps are quite larger than even those, let alone bacterial cells or mold spores that are quite a lot smaller than basidiomycete spores.

edit: Don't let this shy you away from using it if it's what you have! People have and still do use it plenty successfully.

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1diesel1 said:

Is the 12-24 hour soak required before sterilization?
Or can I sterilize after adding all ingredients?

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beluga said:

Missed this.
People do it various ways but what I've found the most effective (for rye) is soaking in tap hot water for about 4 hours in the PC with the lid sealed > boiling that until you see a couple burst grains > then drying very thoroughly > then do your PCing.

There used to be an idea going around that soaking germinates bacteria and spores to help expose them to sterilization, but that was debunked.
The real value is to not hydrate your grain too quickly which leads to lots of burst grain that make your grain a starchy mess.

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Dirtbag said:

@beluga have you used oats before? Thats what i have the easiest access too but i dont see many folks using it, and dont see any tutorial on prepping it. Would it be essentially the same as rye?

Im also looking into millet. Seems a bit fussier to work with but being small it would have a ton of innoculation points compared to a large grain. Way more surface area.

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