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Dnice1
Young Grasshoppa

Registered: 01/13/23
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Millet no prep no soak
#28654219 - 02/10/24 02:06 PM (11 months, 8 hours ago) |
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Hello all,
I recently tried the no soak no prep method of prepping millet for the first time. I put 3.25lbs of millet and 1.75lb of tap water and let it sit in the bag for 12 hours.
Afterwards, I PC’d (let it vent for 15 min) for 2.5 hours. After 4 hours, I took the bag out and sealed it.
After 24 hours, I threw a whole cup of colonized agar in there in front of my laminar flood hood. Unfortunately I didn’t get a picture, but to the eye, it looked clean, uniform growth, no satellite colonies.
After 7 days, the agar had spread onto the grains so I broke it up.
Within 24 hours, I started to spot what I think is trich.
Is this something I’m doing wrong? I’m reading it’s more than likely my spawn. But the steps are so damn easy, I don’t know where I’m messing up.
Note; this was only ONE spawn bag that I tried.
I’m currently prepping my next trial run. I got about 20lbs of millet soaking at the moment that I HAVE rinsed. I’m going to soak for 18 hours, drain for 45 minutes, then load and PC
I have two more bags on the side that I’m going to try and do the NPNS method
I have 6 agar cups that I just took out the fridge that all look the same and came from the same master plate. I’m going to toss them in to the bags… I’ll see what happens
My assumption is if all goes bad - it has to be my agar culture right??
If the NSNP/ or soak goes bad - I’ll have my answer of what works for me…
BUT, what if it’s 1 NSNP, 1-2 prep?! Then id be really confused as to what I’m doing wrong.
Side bar question - should I increase my PC time to over 2.5 hours at 15 psi?
Where I’m at is 164’ over sea level
Thanks to all
Here’s some pictures of what I think is trich on the millet


Edited by Dnice1 (02/10/24 02:15 PM)
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meta_mmxxii


Registered: 08/03/23
Posts: 1,071
Loc: PNW
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Re: Millet no prep no soak [Re: Dnice1] 1
#28654341 - 02/10/24 03:31 PM (11 months, 7 hours ago) |
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First of all, if you are soaking your millet it can hardly be called no soak, it looks to me like you have burst grains I dnt see any trich. the grain looks like it may be over hydrated as well as there is a lot of moisture inside the jar as well. 2 hours is plenty of time for sterilization @15psi at sea level, for all intents and purposes you are. It may be the photos, but I am not seeing any contamination.
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Dnice1
Young Grasshoppa

Registered: 01/13/23
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Quote:
meta_mmxxii said: First of all, if you are soaking your millet it can hardly be called no soak, it looks to me like you have burst grains I dnt see any trich. the grain looks like it may be over hydrated as well as there is a lot of moisture inside the jar as well. 2 hours is plenty of time for sterilization @15psi at sea level, for all intents and purposes you are. It may be the photos, but I am not seeing any contamination.
Hey meta, thanks for the reply.
Here’s another picture of some grain on top that looks like trich. Yes, you are correct. Technically it’s not NPNS, I was just following sandman420s guide as he said that not soaking may lead to the bags stretching and bursting. (I am using bags not jars). I will continue to PC at 2 hours as you suggested. Do you have a millet tek that you follow?


Edited by Dnice1 (02/10/24 04:15 PM)
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Yahra
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Re: Millet no prep no soak [Re: Dnice1] 1
#28654440 - 02/10/24 04:36 PM (11 months, 5 hours ago) |
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Quote:
I threw a whole cup of colonized agar in there
Maybe just use less? Only the best looking, tiny piece of a plate.
The more you put in your bags, the greater the chance of hidden nasties.
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Dnice1
Young Grasshoppa

Registered: 01/13/23
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Re: Millet no prep no soak [Re: Yahra]
#28654493 - 02/10/24 05:25 PM (11 months, 5 hours ago) |
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Quote:
Yahra said:
Quote:
I threw a whole cup of colonized agar in there
Maybe just use less? Only the best looking, tiny piece of a plate.
The more you put in your bags, the greater the chance of hidden nasties.
Hi Yahara. How big of a slice would you recommend? I tried using a small piece in the past and the bag just took so long to finish.
I’m curious. Can trich be enmeshed and hidden in agar for that long and then show itself when it hits grain? Or spawn to bulk?
Also, I should clarify, when I said cup. I meant the ketchup condiment cups
Edited by Dnice1 (02/10/24 05:26 PM)
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HappinessStan
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Re: Millet no prep no soak [Re: Dnice1]
#28654520 - 02/10/24 05:50 PM (11 months, 4 hours ago) |
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It's a shame you don't have photos of the plates or spawn before you shook it up. Not sure about anyone else but, I don't see any mycelium in there. Show us the photos of your new agar cups.
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Dnice1
Young Grasshoppa

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Quote:
HappinessStan said: It's a shame you don't have photos of the plates or spawn before you shook it up. Not sure about anyone else but, I don't see any mycelium in there. Show us the photos of your new agar cups.
Will do. I’ll document this round. In regards to mycelium not showing, I just shook up the bag a day ago, so it should pop back out in a day or so.
Still curious as to the question if trich can stay enmeshed in the agar culture and show after inoculating or spawning to bulk. If not, it should mean at the very least my agar was trich free and it either came from the spawn or maybe a dead spot in the flow hood.
Edited by Dnice1 (02/10/24 11:14 PM)
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Citizen X
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Re: Millet no prep no soak [Re: Dnice1]
#28654880 - 02/10/24 11:45 PM (10 months, 30 days ago) |
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You would also seal the bag first and then PC it 90 minutes. Once your grain comes out of the PC the only thing you want to introduce into that bag is your clean mushroom syringe or agar, but how would you even do that? It’s not like you can open it quickly like a jar
It doesn’t make sense to me how this tek.
How would you introduce agar into a bag
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Edited by Citizen X (02/10/24 11:52 PM)
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Citizen X
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Re: Millet no prep no soak [Re: Citizen X]
#28654887 - 02/10/24 11:55 PM (10 months, 30 days ago) |
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And it looks like 99% of your grains are bursted
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meta_mmxxii


Registered: 08/03/23
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Re: Millet no prep no soak [Re: Citizen X] 2
#28655252 - 02/11/24 10:15 AM (10 months, 30 days ago) |
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Quote:
Citizen X said: You would also seal the bag first and then PC it 90 minutes. Once your grain comes out of the PC the only thing you want to introduce into that bag is your clean mushroom syringe or agar, but how would you even do that? It’s not like you can open it quickly like a jar
It doesn’t make sense to me how this tek.
How would you introduce agar into a bag
Bags get sealed after PC cycle. When ready to inoculate, the bag is then cut open in sterile conditions then add your agar samples and reseal. Hope this clears things up for you.
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LadysKnight
Hello Ladies


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You said you still have some agar from the same master plate that you are going to use on other bags. Got pics?
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Dnice1
Young Grasshoppa

Registered: 01/13/23
Posts: 104
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Quote:
meta_mmxxii said:
Quote:
Citizen X said: You would also seal the bag first and then PC it 90 minutes. Once your grain comes out of the PC the only thing you want to introduce into that bag is your clean mushroom syringe or agar, but how would you even do that? It’s not like you can open it quickly like a jar
It doesn’t make sense to me how this tek.
How would you introduce agar into a bag
Bags get sealed after PC cycle. When ready to inoculate, the bag is then cut open in sterile conditions then add your agar samples and reseal. Hope this clears things up for you.
Hey citizen X,
This is exactly what I do. I fold bags to PC, let it cool down, seal, let is cool down for a day, cut open in front of the flow job, throw in whatever inoculate I’m using, LC, agar etc. and then reseal
Edited by Dnice1 (02/11/24 12:55 PM)
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Dnice1
Young Grasshoppa

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Re: Millet no prep no soak [Re: LadysKnight] 1
#28655475 - 02/11/24 12:49 PM (10 months, 30 days ago) |
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Quote:
LadysKnight said: You said you still have some agar from the same master plate that you are going to use on other bags. Got pics?
Here’s some cups from the same master plate
They just came out of the fridge 2 days ago





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Dnice1
Young Grasshoppa

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Re: Millet no prep no soak [Re: Dnice1]
#28655482 - 02/11/24 12:51 PM (10 months, 30 days ago) |
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I do see some myc today - but wanted to see if someone can confirm that’s trich. I’m waiting to see if it spreads before I toss, but if someone can ID it early, it’s going to the trash.

Edited by Dnice1 (02/11/24 12:58 PM)
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LadysKnight
Hello Ladies


Registered: 10/09/15
Posts: 2,331
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Re: Millet no prep no soak [Re: Dnice1]
#28655542 - 02/11/24 01:36 PM (10 months, 30 days ago) |
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I mean... Trich or not, it's green. So, it's fucked. While those plates are showing mostly mushroom mycelium, they are not clean. Look on the edges with the least growth, you will see a lot of disembodied fuzz.
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Dnice1
Young Grasshoppa

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Quote:
LadysKnight said: I mean... Trich or not, it's green. So, it's fucked. While those plates are showing mostly mushroom mycelium, they are not clean. Look on the edges with the least growth, you will see a lot of disembodied fuzz.
Quote:
LadysKnight said: I mean... Trich or not, it's green. So, it's fucked. While those plates are showing mostly mushroom mycelium, they are not clean. Look on the edges with the least growth, you will see a lot of disembodied fuzz.
Got it. So you’re thinking my cultures bad. More transfers? It’s about 4 transfers out from a clone
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LadysKnight
Hello Ladies


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Posts: 2,331
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Re: Millet no prep no soak [Re: Dnice1]
#28655598 - 02/11/24 02:39 PM (10 months, 30 days ago) |
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Yes, sure continue to try to clean that up, it will be good practice. But meanwhile start something new from different sources if possible.
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Dnice1
Young Grasshoppa

Registered: 01/13/23
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Well, I’m already an hour into my PC with grains and it’ll take me a while to clean up the culture - I do have some proven LC in the fridge I can use. So we’ll see how that goes.
Here’s some pics of millet that was rinsed and soaked for 18 hours Vs Millet soaked in bags for 12 hours without rinse.
I’m thinking about cutting an Agar cup in half throwing half in one each.
If the cultures bad, both bags should contam. We’ll see how it turns out

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LadysKnight
Hello Ladies


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Re: Millet no prep no soak [Re: Dnice1]
#28655657 - 02/11/24 03:25 PM (10 months, 30 days ago) |
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I like experiments, moor data!
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Shaw


Registered: 06/27/01
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Re: Millet no prep no soak [Re: Dnice1] 1
#28655947 - 02/11/24 06:49 PM (10 months, 30 days ago) |
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I read years ago, (maybe TMC) that you didn't want to even use agar plates that were fully colonized. Just a colonized piece of it. The reasoning here is that if a plate got contaminated it would be less noticeable without uncolonized agar for it to show up on.
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