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Doi
Stranger
Registered: 06/03/16
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Last seen: 3 years, 7 months
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For anyone who has the time
#23436573 - 07/12/16 11:49 PM (7 years, 6 months ago) |
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Hey all,
I've started the cultivation process of p subs using mycelium found from bark under the patch and placing on agar, the strongest myc was then isolated. The same procedure was done again but using a stem from a p sub pin placed on agar.
Here are some pictures of the agar plates with wedges taken out:




I have taken three agar wedges, one for each jar. Whole rye grain was used (100g for each 500ml jar mixed with 100-110g of water. Each jar was fitted with a polyfil filter and pressure cooked for one hour. Jars were taken out and shaken to separate wet from dry grain. Once jars were completely cooled agar wedges placed on-top of rye grain and left still.
Here are some pictures of the jars:


I am posting this as some may find it interesting and I wish to know if I am on track or if any crucial mistakes have been made.
Thanks!
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Psilosoulful

Registered: 09/05/14
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Re: For anyone who has the time [Re: Doi]
#23436735 - 07/13/16 01:00 AM (7 years, 6 months ago) |
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Not bad, hopefully those grains dry up a bit more over the next few days, they're a bit on the wet side. And try to pressure cook your grain jars for at least 90 minutes. You want to kill off as many endospores as possible.
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Doi
Stranger
Registered: 06/03/16
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Cheers, I used this guide: http://www.fungifun.org/English/Grain-For-Simple-Minds
Does it matter where the jars are stored while the mycelium builds up? They obviously require gas exchange, but should they be placed in an extremely sterile area? or is it okay to leave them in a drawer or something.
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Bunya

Registered: 06/04/16
Posts: 601
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Re: For anyone who has the time [Re: Doi]
#23436830 - 07/13/16 02:16 AM (7 years, 6 months ago) |
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Nice job.
-------------------- Bunya cones kill.
Edited by Bunya (01/04/17 12:12 AM)
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Bunya

Registered: 06/04/16
Posts: 601
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Re: For anyone who has the time [Re: Bunya]
#23436834 - 07/13/16 02:19 AM (7 years, 6 months ago) |
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Also have you tried a liquid culture with the agar?
-------------------- Bunya cones kill.
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NDStepp84
Stellar nuclear waste


Registered: 04/23/15
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Re: For anyone who has the time [Re: Doi]
#23436939 - 07/13/16 04:33 AM (7 years, 6 months ago) |
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Do you have extremely sterile areas in your house? I know I don't. A shelf or something is fine.
--------------------
"I am free, no matter what rules surround me. If I find them tolerable, I tolerate them; if I find them too obnoxious, I break them. I am free because I know that I alone am morally responsible for everything I do. -Robert A. Heinlein
Links and teks ND's grow log and discussion Plant thread
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Inocuole
Scalpel of Evil's Bane



Registered: 11/21/11
Posts: 24,863
Loc: ★
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Re: For anyone who has the time [Re: NDStepp84]
#23436954 - 07/13/16 04:46 AM (7 years, 6 months ago) |
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Those grains look so bacterial. I wouldn't even put agar to them, straight to the trash.
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weetsie
unlicensed tub surgeon



Registered: 05/08/11
Posts: 572
Loc: United Kingdom
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Re: For anyone who has the time [Re: Inocuole]
#23436967 - 07/13/16 04:53 AM (7 years, 6 months ago) |
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Quote:
Inocuole said: Those grains look so bacterial. I wouldn't even put agar to them, straight to the trash.
That's just what they look like when you dont cook them first, especially if you use 110% water.
They can't be bacterial if they only came out of the PC a few hours before the photo was taken.
-------------------- Active grow logs: Oysters on Straw Pellets Trade list
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Inocuole
Scalpel of Evil's Bane



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Re: For anyone who has the time [Re: weetsie]
#23437044 - 07/13/16 06:08 AM (7 years, 6 months ago) |
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Quote:
weetsie said: They can't be bacterial if they only came out of the PC a few hours before the photo was taken.
Wanna bet?
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Mad Season
hookers and blackjack



Registered: 09/16/12
Posts: 12,666
Loc: Canada
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Re: For anyone who has the time [Re: Inocuole]
#23437057 - 07/13/16 06:14 AM (7 years, 6 months ago) |
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They may be just hella starchy RN but there are endospores right now just waiting to germinate. Once they do, they'll just rip through the starches. Yeah that shit is going to end up being bacterial af.
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Mycologist217
Frank's Disciple



Registered: 02/13/13
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Re: For anyone who has the time [Re: Mad Season]
#23437253 - 07/13/16 08:03 AM (7 years, 6 months ago) |
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I've been using a 65 minute 15PSI pressure sterilization for my birdseed for probably my last 100 jars, and have not had any issues.
-------------------- My LC Manual (With custom LC lid Tek) ~~ Required Mycology Supplies ~~ Agar Work Videos ~~ L G M AMU Q&A-NO SYMPATHY FOR THE DEVIL! KEEP THAT IN MIND! BUY THE TICKET: TAKE THE RIDE Check out my Retail Gourmet Mushroom Farm! Mycologist217 is a fictitious entity that uses images supplied by Google to mask his/her inability to develop normal social habits.
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Mad Season
hookers and blackjack



Registered: 09/16/12
Posts: 12,666
Loc: Canada
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I think we're talking about how it's sticky and starchy, which is more a prep problem. 65 minutes is a little low though. After changing up to 120 minutes with wbs I was having MUCH less wet grains.
Quote:
Mad Season said: Oh dear lord no. I'll get a quote from when I had this debate awhile back. There's a big difference between fully colonized and cleanly colonized.
Quote:
Mad Season said: Well there's a difference between colonized and cleanly colonized.
This is colonized: 
Lots of members have noticed cobweb and trich from jars that look like this or better. Wet spots are bacteria. Not wet grains or wet spots. To colonize cleanly is the hardest. I had to change my pc times to 2 hours to reduce it.
This is (ALMOST) cleanly colonized, that's how hard it is to have it completely clean. This isn't even completely clean:

This is completely clean spawn:

Anyone who's done this for awhile will testify this is the hardest part of the hobby and is what will dictate the success or failure of bulk.
All those jars had a mushroomy smell. A miniscule amount of bacteria will get overpowered by the mushroom musk. Especially if it is covering the surface
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weetsie
unlicensed tub surgeon



Registered: 05/08/11
Posts: 572
Loc: United Kingdom
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Re: For anyone who has the time [Re: Inocuole]
#23437269 - 07/13/16 08:11 AM (7 years, 6 months ago) |
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Quote:
Inocuole said:
Quote:
weetsie said: They can't be bacterial if they only came out of the PC a few hours before the photo was taken.
Wanna bet?
OK technically they could be but what I'm saying is they look that way because of how the grain was prepped, there is no viable contam in those photos.
-------------------- Active grow logs: Oysters on Straw Pellets Trade list
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Inocuole
Scalpel of Evil's Bane



Registered: 11/21/11
Posts: 24,863
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Re: For anyone who has the time [Re: weetsie]
#23437275 - 07/13/16 08:13 AM (7 years, 6 months ago) |
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There are, they'll be waking up to a moist delicious feast soon by the looks of it.
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Mycologist217
Frank's Disciple



Registered: 02/13/13
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Re: For anyone who has the time [Re: Inocuole]
#23437294 - 07/13/16 08:23 AM (7 years, 6 months ago) |
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Oh yeah....hahaha I was pretty worried when I first started getting that whispy colonization you have there in the completely clean pic, but Inocuole was the one that told me it is indicative of exceptionally clean growth, now it's what I aim for:
 Sorry my camera is so snotty...hope you can see the resemblance to your example!
As opposed to this:

-------------------- My LC Manual (With custom LC lid Tek) ~~ Required Mycology Supplies ~~ Agar Work Videos ~~ L G M AMU Q&A-NO SYMPATHY FOR THE DEVIL! KEEP THAT IN MIND! BUY THE TICKET: TAKE THE RIDE Check out my Retail Gourmet Mushroom Farm! Mycologist217 is a fictitious entity that uses images supplied by Google to mask his/her inability to develop normal social habits.
Edited by Mycologist217 (07/13/16 08:33 AM)
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Doi
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Registered: 06/03/16
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Re: For anyone who has the time [Re: Doi]
#23439878 - 07/14/16 03:49 AM (7 years, 6 months ago) |
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So after looking through the comments I decided to do another jar. I brang 200g of water to the boil and placed 100g rye into this. The heat was then turned to a simmer and the rye was stirred occasionally, after 30 minutes there was no water left and the rye started sticking to the bottom, I added maybe another ~30ml of water to it and simmered for another 10 min. After simmering the rye was strained and a little cold water was poured through it to remove starch. The rye was then placed in a jar and pressure cooked for 70 minutes.
This is the result:

It looks a lot cleaner, although still very wet. There is no starch stuck to the sides of the jar and the rye seems to be quite loose / not sticky.
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PsilocyBen17
Pin Pornographer


Registered: 10/20/13
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Re: For anyone who has the time [Re: Doi]
#23440274 - 07/14/16 08:52 AM (7 years, 6 months ago) |
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follow a proper grain prep tek. I don't like boiling rye. too many burst grains. For rye I prefer to soak overnight, simmer, drain, pc.
Oats I don't mind boiling if I forget to soak them the night before. They are so damn tough that they are impossible to fuck up.
And, as I learned from Innocule, they bounce back much faster after a shake or g2g then rye does. I guess this is attributed to the hard hull(s)
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weetsie
unlicensed tub surgeon



Registered: 05/08/11
Posts: 572
Loc: United Kingdom
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If you're cooking the grain before hand just use plenty of water and stain off the excess once it's done.
-------------------- Active grow logs: Oysters on Straw Pellets Trade list
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Doi
Stranger
Registered: 06/03/16
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Last seen: 3 years, 7 months
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Re: For anyone who has the time [Re: weetsie]
#23446093 - 07/16/16 07:23 AM (7 years, 6 months ago) |
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Should I expect to be seeing any growth in the grain at this point? Agar was put in on 12/7 10PM
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weetsie
unlicensed tub surgeon



Registered: 05/08/11
Posts: 572
Loc: United Kingdom
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Re: For anyone who has the time [Re: Doi]
#23446144 - 07/16/16 07:58 AM (7 years, 6 months ago) |
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Normally the wedge will fluff up by the next day, within a few days even a slow species should be colonizing the grain surrounding the wedge.
-------------------- Active grow logs: Oysters on Straw Pellets Trade list
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Doi
Stranger
Registered: 06/03/16
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Re: For anyone who has the time [Re: weetsie]
#23446264 - 07/16/16 09:02 AM (7 years, 6 months ago) |
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I'm not even sure if the myc will colonize the grain at this point. Seems to be 0 growth.
Threads such as https://www.shroomery.org/forums/showflat.php/Number/18022898#18022898 have users saying that agar to grain for this species hasn't worked 
Idk how to cultivate P. Subaeruginosa at this point. Might try relocating soil already mixed with myc and chips onto an outdoor patch with cardboard.
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Mad Season
hookers and blackjack



Registered: 09/16/12
Posts: 12,666
Loc: Canada
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Re: For anyone who has the time [Re: Doi]
#23446280 - 07/16/16 09:13 AM (7 years, 6 months ago) |
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I have never had an issue with P.subs and agar to grain. There's something else going on if that's the case. Like the grain is all mushy, wet, starchy, and pretty damn funky looking. Just inviting bacteria.
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Doi
Stranger
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Re: For anyone who has the time [Re: Mad Season]
#23447611 - 07/16/16 07:04 PM (7 years, 6 months ago) |
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My last jar is completely clean, no starch, stickiness and it seems to be pretty dry at this point. The agar wedges aren't fluffing up at all, idk if this is a characteristic of myc on agar and this never happened in the petri dishes, starting to think that the mycelium growth in agar has gone wrong or something.
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just_curious
Cultivator


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Re: For anyone who has the time [Re: Doi]
#23447842 - 07/16/16 08:56 PM (7 years, 6 months ago) |
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-I add 1 1/4 cup of grain for every quart jar I plan on using
-I put it all in my pressure cooker (because it can hold all the grains)
- I got my tap water extremely hot, and I rinse the grains about 5 times. Being sure to dunk all the dirty water before the next rinse.
- on the 5th rinse, I leave in the water, add 1 cup of liquid coffee from my mornings pot, 2 tablespoons of gypsum, and then soak for roughly 24 hours.
- at the 24 hour mark, I place the PC on the stove, lid on but not locked in so it can't pressure cook, I turn the stove on high, and let it come to a boil.
- once it is at the boil, I start my 20 minute timer. I stir it maybe one or twice during the 20 minutes but don't adjust the heat.
- I place 2 strainers in my sink, and after 20 minutes, I dump the grains in equal amounts into the strainers. At this point I grab a large spoon and stir it so the steam evaporates all the water off the grains. (I noticed when you shake the strainers, the grains fall through or fall out. Also, this is why I do 1 1/4 cup instead of 1 cup for every quart)
-once I get it to the point where I'm not seeing steam any more, I dump the grains onto a large, flat, Deep tray so more surface area is exposed. At this point the last little bit of cooling can happen.
- once my grains are completely cooled to room temperature, I add the grains to my jars. (I noticed when you put them in with the grains still hot, that steam is still going and that evaporation has nowhere to go, ultimately building up condensation on the inside of the jars, causing wet grains inside the jar)
- when they're loaded up, I PC. I have a weight in my PC instead of a gauge. I put the heat on high until I hear the load steam come out, and then I drop the heat between 6 and 7 and set the timer for 90 minutes.
I don't get bursted grains, or wet grains this way. They always look dry in appearance, but are wet on the inside. The gypsum makes it so the don't stick together either. At no point do I rinse my grains after the boil. Really all the boiling is for is to assist with evaporating all the water off. I started this using RR's grain prep, but added in a couple steps that I felt benefited me. But without his prep TEK, I would've been lost. Trying to evaporate and cool the grains in the strainer alone takes FOREVER!!
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