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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,819
Loc: Canada
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Re: Cultivation General Discussion [Re: Steevo] 1
#24652501 - 09/22/17 08:12 PM (6 years, 5 months ago) |
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If I left no pour plates in a PC for 2 weeks I wouldn't even bother to resterilize. Just pull em out and use em. Unless you forgot to PC them to begin with. Then I would just put em in the trash.
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tryptkaloids
Learner



Registered: 02/08/15
Posts: 12,649
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Re: Cultivation General Discussion [Re: Pastywhyte]
#24652511 - 09/22/17 08:17 PM (6 years, 5 months ago) |
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if you have properly sterilized them they should never grow anything unless opened, they should dry out long before anything has a chance
-------------------- "Remember, kids, the difference between science and screwing around is writing it down" -adam savage Flowchart for Recommended plan of action. Learn the tried and true way to grow mushrooms Use the Damn search engine After you know what you're doing, take a break Pick a book, Make some chips! Josex said:Don't take the site seriously bro, ain't worth it.
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Josex
#cheat_code


Registered: 11/13/15
Posts: 8,995
Loc:
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Re: Cultivation General Discussion [Re: natedawgnow]
#24652578 - 09/22/17 08:54 PM (6 years, 5 months ago) |
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Quote:
natedawgnow said: I've read it too but I don't know it takes some high temps to caramelize malt and glucose. Fructose will fail at 220 or so but malt is a strong complex sugar. As is dextrose. Maybe at extended times but I don't know man I still don't think 45min at anything less than 300 degrees is enough to do in malt or dextrose.
But I could very well be wrong.
I recently pc'd no-pour containers with LME agar at 20 psi for 35 min and had to trash it all and start all over, got god awful caramelization, didn't want to take my chances after having read that the Maillard reaction may fuck up your cultures. Gotta pc LME at 15 psi. As far as extended times is concerned, I've pc'd LME for as long as 50 mins at 15 psi and got no caramelization whatsoever. My gut tells me pc times have very little to do with caramelization ocurring, I think you could go beyond one hour at 15 psi without getting any.
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Munchauzen


Registered: 06/22/11
Posts: 14,343
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Re: Cultivation General Discussion [Re: Pastywhyte] 1
#24652592 - 09/22/17 08:59 PM (6 years, 5 months ago) |
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Quote:
Pastywhyte said: If I left no pour plates in a PC for 2 weeks I wouldn't even bother to resterilize. Just pull em out and use em. Unless you forgot to PC them to begin with. Then I would just put em in the trash.
I once used a slant that was over a year old since sterilization before I nocced it up. worked fine. transfered that slant a few months ago to a new one.
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natedawgnow
Rocky mountain hood rat



Registered: 02/09/15
Posts: 8,940
Loc: ation
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Re: Cultivation General Discussion [Re: Munchauzen]
#24652607 - 09/22/17 09:07 PM (6 years, 5 months ago) |
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Even at 20psi you still aren't reaching the 350 degrees F it takes to caramelize malt.
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Josex
#cheat_code


Registered: 11/13/15
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Re: Cultivation General Discussion [Re: natedawgnow]
#24652629 - 09/22/17 09:20 PM (6 years, 5 months ago) |
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Yeah I know, it's a much lower temp All I know I had to trash a whole batch cos my shit caramelized. Try it if you want just for science, I ain't making nothing up.
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herrenvolk


Registered: 05/14/16
Posts: 222
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Re: Cultivation General Discussion [Re: Josex]
#24652686 - 09/22/17 09:57 PM (6 years, 5 months ago) |
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I've PC'ed PDA agar for 2 hours at 15psi. The agar comes out darker than usual and growth seems to be somewhat slower than usual. But still usable.
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herrenvolk


Registered: 05/14/16
Posts: 222
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Re: Cultivation General Discussion [Re: herrenvolk]
#24652894 - 09/23/17 12:52 AM (6 years, 5 months ago) |
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I've got a few questions for you folks that use a magnetic stir bar for stirring LCs. 1. How often do you stir the lcs? Will stirring often speed up growth? Will stirring too often inhibit growth? 2. How long do you let it stir, and how fast? Is it possible to harm the culture by stirring too hard/too long?
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natedawgnow
Rocky mountain hood rat



Registered: 02/09/15
Posts: 8,940
Loc: ation
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Re: Cultivation General Discussion [Re: herrenvolk]
#24652943 - 09/23/17 02:14 AM (6 years, 5 months ago) |
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Nah I definitely don't think you are making something up but are you for sure they actually caramelized? Like they turned brown and formed a thicker viscosity and opaqueness? Did you taste it by chance I will definitely do a run for science. I almost always do my agar at 17 or so psi for 30 min. I have a weight for 20 so I'll do it and observe the difference myself.
What kind of malt do you use? Maybe brewers malt extract is less pure allowing for lower caramelization temps 
Pure maltose shouldn't degrade at all till well over 300 degrees F but I can see a complex chain of impurities causing lower caramelization temps. If you're using pda and use actual dextrose I wouldn't worry about it at all
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Josex
#cheat_code


Registered: 11/13/15
Posts: 8,995
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Re: Cultivation General Discussion [Re: natedawgnow]
#24652962 - 09/23/17 02:59 AM (6 years, 5 months ago) |
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Pretty sure man, I dye my agar blue and when I looked at the bottom of the plates after they solidified I saw lots of little brownish dots standing out from the blue, never got to taste the agar though. I use spraymalt, extra light for brewers, it says that in the label. It's a fine powder that absorbs the humidity of the air like a motherfucker, isn't that what you use then? Also I don't know if sterilizing in media bottles will make a difference, I do no pours.
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Mateja



Registered: 07/14/16
Posts: 7,948
Loc: Here
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Re: Cultivation General Discussion [Re: Josex]
#24653149 - 09/23/17 07:15 AM (6 years, 5 months ago) |
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Is 45 min PC enough for WBS agar?
I boil 30g WBS in 3dl water for 15 minutes, after that i throw away the sseds and mix the broth with agar-agar and throw in the PC. I have read that for liquid is 45 min PC, but the endospores are still in this water mix right? And they are sufficiently beat up in 45 min?
-------------------- Cakes inside Water Tub
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tryptkaloids
Learner



Registered: 02/08/15
Posts: 12,649
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Last seen: 11 hours, 26 minutes
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Re: Cultivation General Discussion [Re: Mateja]
#24653159 - 09/23/17 07:22 AM (6 years, 5 months ago) |
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any liquids should be effectively sterilized in that time at 15psi. certain media like dog food and pda have high populations in them and need that long. for other things I usually only go 20-30 mins
-------------------- "Remember, kids, the difference between science and screwing around is writing it down" -adam savage Flowchart for Recommended plan of action. Learn the tried and true way to grow mushrooms Use the Damn search engine After you know what you're doing, take a break Pick a book, Make some chips! Josex said:Don't take the site seriously bro, ain't worth it.
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Mateja



Registered: 07/14/16
Posts: 7,948
Loc: Here
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Re: Cultivation General Discussion [Re: tryptkaloids]
#24653165 - 09/23/17 07:29 AM (6 years, 5 months ago) |
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I thought I was ballin for a while pouring my DFA plates, all clean, until i started pouring WBS agar, ALL contamed at pouring. I'm PCing second batch WBS agar now and I will go out of my ways to try to pour them as clean as i can and we'll see, wish me luck.
-------------------- Cakes inside Water Tub
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tryptkaloids
Learner



Registered: 02/08/15
Posts: 12,649
Loc: Exact Center
Last seen: 11 hours, 26 minutes
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Re: Cultivation General Discussion [Re: Mateja]
#24653189 - 09/23/17 07:47 AM (6 years, 5 months ago) |
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hmmm. you sure its not technique?
-------------------- "Remember, kids, the difference between science and screwing around is writing it down" -adam savage Flowchart for Recommended plan of action. Learn the tried and true way to grow mushrooms Use the Damn search engine After you know what you're doing, take a break Pick a book, Make some chips! Josex said:Don't take the site seriously bro, ain't worth it.
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,890
Loc: Milky way
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Re: Cultivation General Discussion [Re: tryptkaloids]
#24653196 - 09/23/17 07:55 AM (6 years, 5 months ago) |
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They make autoclave test strips. Loaded with over one million of the most resistant endospores. And in a volume of liquid under 1 liter it only takes 15m at 15psi to inactivate all of them a 1/10,000,000 probability of a single survivor.
Quote:
Biological autoclave indicators
Another form of autoclave indicator makes use of biological indicators. These are standardized populations of resistant bacterial spores such as Geobacillus stearothermophilus. This bacterium comes in spore form on strips or as a suspension in prepackaged vials. This bacterial spore test is used to determine if the sterilization cycle parameters were sufficient to kill the test microorganisms. This biological indicator of sterilization success is a good one and should be done every time you autoclave. It is best to place the biological indicator in an item to be autoclaved and best if placed at the center of the load.
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Mateja



Registered: 07/14/16
Posts: 7,948
Loc: Here
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Re: Cultivation General Discussion [Re: tryptkaloids]
#24653318 - 09/23/17 09:12 AM (6 years, 5 months ago) |
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Quote:
tryptkaloids said: hmmm. you sure its not technique?
First I thought it was my transfer-technique but now I understand it was my pouring-technique that was fault. None of my DFA plates showed contam at pouring nor after transferring, but all WBS plates were contained at pouring it seems (I just took over 4 days for the contains to germinate)
New bottle WBS is cooling now, so in one hour I'll try again. And I will wait 4 days before making transfers.
-------------------- Cakes inside Water Tub
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,819
Loc: Canada
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Re: Cultivation General Discussion [Re: Mateja]
#24653336 - 09/23/17 09:22 AM (6 years, 5 months ago) |
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I know the schools of thought state that 15 min is sufficient. But I have had blank plates go totally bacterial on me from trying shorter runs. I am of the belief that an extra 30 min is worth not having to lose all my plates 3 days later.
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tryptkaloids
Learner



Registered: 02/08/15
Posts: 12,649
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Re: Cultivation General Discussion [Re: Pastywhyte]
#24653354 - 09/23/17 09:35 AM (6 years, 5 months ago) |
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what elevation are you at?
-------------------- "Remember, kids, the difference between science and screwing around is writing it down" -adam savage Flowchart for Recommended plan of action. Learn the tried and true way to grow mushrooms Use the Damn search engine After you know what you're doing, take a break Pick a book, Make some chips! Josex said:Don't take the site seriously bro, ain't worth it.
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,890
Loc: Milky way
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Re: Cultivation General Discussion [Re: tryptkaloids]
#24653411 - 09/23/17 10:04 AM (6 years, 5 months ago) |
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Improper venting is the no1 cause of insufficient sterilization. I think its actually why we have to do grains so long because the air never actually fully gets out even if you vent 20-30m. Just a hunch on that tho
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,819
Loc: Canada
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Re: Cultivation General Discussion [Re: bodhisatta]
#24653448 - 09/23/17 10:16 AM (6 years, 5 months ago) |
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I'm pretty sure I know how to vent my sterilizer. My elevation is well over 2000 ft.
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