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OfflineTEAMSESH
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germination problem
    #22053427 - 08/06/15 12:30 PM (8 years, 5 months ago)

I get no mysilum and it just contams on agar
print is one year old
I'm using MEA for germination.

need help .


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Offlinespacechildo
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Re: germination problem [Re: TEAMSESH]
    #22054162 - 08/06/15 03:36 PM (8 years, 5 months ago)

you must have a really dirty print or do your agar completely wrong.

do your pour agar into plates in a SAB or do you PC small containers with agar already in them?

have you tried just leaving some plates without inoculating and see if they also contaminate?

try using less spores, and especially less spore solution if you are using a syringe, 1 drop max.


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OfflineTEAMSESH
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Re: germination problem [Re: spacechildo]
    #22054898 - 08/06/15 06:26 PM (8 years, 5 months ago)

might be my sterilization .
im sure I did the agar right on sterilized plates.    and I used a STILL AIR box blasted with 99.9 germ bacteria virus spray you can buy at Walmart. ill try again this week. only have half a print left and living in socal its tuff to come by prints pm for more help if you can.


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OfflineAppalachian Brony
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Re: germination problem [Re: TEAMSESH]
    #22054931 - 08/06/15 06:30 PM (8 years, 5 months ago)

Why do you have two identical threads, spelling germination differently, about a ywo year old and one year old print? You could probably cover both old prints in one thread.


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OfflineTEAMSESH
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Re: germination problem [Re: Appalachian Brony]
    #22054941 - 08/06/15 06:32 PM (8 years, 5 months ago)

sorry its actually 2 years old now


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InvisibleBuck513
Registered: 04/17/14
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Re: germination problem [Re: Appalachian Brony]
    #22054947 - 08/06/15 06:33 PM (8 years, 5 months ago)

What you spray inside your SAB isn't nearly as important as how you move inside it. Most people use soapy water on plain old water in their SAB.

I wouldn't even waste money on that spray. Learn how to move inside a SAB


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OfflineAppalachian Brony
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Re: germination problem [Re: TEAMSESH]
    #22054956 - 08/06/15 06:34 PM (8 years, 5 months ago)

Time flies, check your other thread tho bro


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OfflineTEAMSESH
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Re: germination problem [Re: Buck513]
    #22054990 - 08/06/15 06:40 PM (8 years, 5 months ago)

well I use doctors gloves, iso 90% 

food grade agar and light malt extract. 
I pressure cooked that for like 35 mins at some psi idk its a 16 qrt from Walmart.


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InvisibleBuck513
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Re: germination problem [Re: TEAMSESH]
    #22054998 - 08/06/15 06:41 PM (8 years, 5 months ago)

70% Iso is better, and 35 mins might be enough but you should bump it up to 45 or so


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InvisiblebodhisattaMDiscordReddit
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Re: germination problem [Re: Buck513]
    #22055026 - 08/06/15 06:50 PM (8 years, 5 months ago)

agar only needs to be PC for 15-20m.

and 70% is far superior as a sanitizer than 91% you can turn 91% into 70% with distilled water

(70%)(130ml)=(91%)(100ml)
so to 100ml of 91% add 30ml H2O to achieve 70%


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OfflineTEAMSESH
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Re: germination problem [Re: Buck513]
    #22055030 - 08/06/15 06:50 PM (8 years, 5 months ago)

12-45 mins does it really make a deferens ? more the better ?

pre sterilized plates an gling wrap was used cut about a 2 inch off the whole roll so its be easy to wrap the dish.'s


Edited by TEAMSESH (08/06/15 06:52 PM)


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OfflineAppalachian Brony
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Re: germination problem [Re: TEAMSESH]
    #22055053 - 08/06/15 06:57 PM (8 years, 5 months ago)

If the plates were pre-sterilized there was no reason to sterilize them


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OfflinePsilosoulful

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Re: germination problem [Re: bodhisatta]
    #22055131 - 08/06/15 07:13 PM (8 years, 5 months ago)

Quote:

bodhisatta said:
agar only needs to be PC for 15-20m.



:orry:


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InvisiblebodhisattaMDiscordReddit
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Re: germination problem [Re: Psilosoulful]
    #22055181 - 08/06/15 07:25 PM (8 years, 5 months ago)



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InvisiblebodhisattaMDiscordReddit
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Re: germination problem [Re: bodhisatta]
    #22055191 - 08/06/15 07:29 PM (8 years, 5 months ago)

agar is liquid it sterilizes very easily.
also when canning you fill with liquid til you cover the item being canned for this same reason.
grains we don't cover in water so they take a while longer.

spore test evaluations use the hardest to kill endospores and are placed in a bottle of agar. it goes through the sterilization cycle and then the endospore strip is incubated on a special agar in a temperature controlled incubator. 5-7 days later if you get no growth you get a confirmation of effective sterilization. they usually have 1x10^7 or more spores on the test strips to make sure you achieve at least 7 logs of kill. if you can kill the very hardest endospores in 15m in agar I would say 20+ is overkill.

Geobacillus stearothermophilus
https://en.wikipedia.org/wiki/Bacillus_stearothermophilus
Quote:

It is commonly used as a challenge organism for sterilization validation studies and periodic check of sterilization cycles. The biological indicator contains spores of the organism on filter paper inside a vial. After sterilizing, the cap is closed, an ampule of growth medium inside of the vial is crushed and the whole vial is incubated. A color and/or turbidity change indicates the results of the sterilization process; no change indicates that the sterilization conditions were achieved, otherwise the growth of the spores indicates that the sterilization process has not been met.

Biological indicators are used in conjunction with chemical indicators and process indicators to validate sterilization processes





you can buy chemical test strips but the biological indicators are accurate. the chemical strips are more of a indicator that a cycle has been ran or not depending on the color of the indicator.

biological indicators require a special incubator and media. out of reach for most cube cultivators.


Edited by Trusted cuItivator (08/06/15 07:32 PM)


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Offlinekushroom
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Re: germination problem [Re: TEAMSESH]
    #22055232 - 08/06/15 07:40 PM (8 years, 5 months ago)

Quote:

TEAMSESH said:
well I use doctors gloves, iso 90% 

food grade agar and light malt extract. 
I pressure cooked that for like 35 mins at some psi idk its a 16 qrt from Walmart.




Let me guess your rolling with the presto one? I got mine from walmart too it was like $70


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InvisibleMad Season
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Re: germination problem [Re: kushroom]
    #22055270 - 08/06/15 07:51 PM (8 years, 5 months ago)



--------------------
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OfflinePsilosoulful

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Re: germination problem [Re: bodhisatta]
    #22055388 - 08/06/15 08:16 PM (8 years, 5 months ago)

Quote:

bodhisatta said:
agar is liquid it sterilizes very easily.
also when canning you fill with liquid til you cover the item being canned for this same reason.
grains we don't cover in water so they take a while longer.

spore test evaluations use the hardest to kill endospores and are placed in a bottle of agar. it goes through the sterilization cycle and then the endospore strip is incubated on a special agar in a temperature controlled incubator. 5-7 days later if you get no growth you get a confirmation of effective sterilization. they usually have 1x10^7 or more spores on the test strips to make sure you achieve at least 7 logs of kill. if you can kill the very hardest endospores in 15m in agar I would say 20+ is overkill.

Geobacillus stearothermophilus
https://en.wikipedia.org/wiki/Bacillus_stearothermophilus
Quote:

It is commonly used as a challenge organism for sterilization validation studies and periodic check of sterilization cycles. The biological indicator contains spores of the organism on filter paper inside a vial. After sterilizing, the cap is closed, an ampule of growth medium inside of the vial is crushed and the whole vial is incubated. A color and/or turbidity change indicates the results of the sterilization process; no change indicates that the sterilization conditions were achieved, otherwise the growth of the spores indicates that the sterilization process has not been met.

Biological indicators are used in conjunction with chemical indicators and process indicators to validate sterilization processes





you can buy chemical test strips but the biological indicators are accurate. the chemical strips are more of a indicator that a cycle has been ran or not depending on the color of the indicator.

biological indicators require a special incubator and media. out of reach for most cube cultivators.



Interesting info, but I'm not gonna use poured agar, I'll be using "pasty" plates instead. Pasty recommends 45 min, so I guess that type of agar will take longer to sterilize completely, being as you cannot place the mini round directly on the bottom of the canner, it has to be propped up by jar lids or quart jars.


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InvisibleMad Season
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Re: germination problem [Re: Psilosoulful]
    #22055417 - 08/06/15 08:22 PM (8 years, 5 months ago)

If anything pasty plates get sterilized quicker. Less liquid means it'll heat up and get to temperature much quicker. Liquids consistency and its amazing ability of convection makes it heat up much quicker than grains


--------------------
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How to shroomery like a pro! (Seriously, everyone read this!)
Improve your sterile techniques! (A comprehensive guide to agar)
Links upon links of literally EVERYTHING UP TO DATE

AMU Q&A
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InvisiblebodhisattaMDiscordReddit
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Re: germination problem [Re: Psilosoulful]
    #22055423 - 08/06/15 08:23 PM (8 years, 5 months ago)

doesn't matter it's the same agar. I know pasty says 45 but 15 would do it too. it doesn't matter you don't want your agar bottle on the bottom of the PC either.

I've never sterilized my home made or pre mix agar longer than 20m. and also never had a problem with the agar itself,

your PC takes longer to get to pressure when full, it doesn't get to 15 until things are up to temp on the inside, it's a property of the latent heat transfer of steam, it condenses on the cooler jars and looses a insane amount of heat energy until the jars heat up it won't get to 15psi.
you start the timer at 15psi.

regardless of how much agar is in your PC it will still take 15m in a 500ml jar or several small pasty plates.


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OfflinePsilosoulful

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Re: germination problem [Re: bodhisatta]
    #22055443 - 08/06/15 08:29 PM (8 years, 5 months ago)

Quote:

bodhisatta said:
doesn't matter it's the same agar. I know pasty says 45 but 15 would do it too. it doesn't matter you don't want your agar bottle on the bottom of the PC either.

I've never sterilized my home made or pre mix agar longer than 20m. and also never had a problem with the agar itself,

your PC takes longer to get to pressure when full, it doesn't get to 15 until things are up to temp on the inside, it's a property of the latent heat transfer of steam, it condenses on the cooler jars and looses a insane amount of heat energy until the jars heat up it won't get to 15psi.
you start the timer at 15psi.

regardless of how much agar is in your PC it will still take 15m in a 500ml jar or several small pasty plates.



Thanks for the info bod, I noticed other cultivators complaining of too much condensation in their plates, agar being too stiff, etc. It is probably because PC times are too long.

And also, when I made my agar substitute tek (brf and water), I PC'd for 15 min at 15 PC and all my brf paste jars came out good with only one contaminant out of 6 jars (probably due to lack of proper sterile technique), still a success IMO.


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InvisibleMad Season
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Re: germination problem [Re: Psilosoulful]
    #22055457 - 08/06/15 08:32 PM (8 years, 5 months ago)

Quote:

MA - Care must be taken used with malt not to exceed 10 pounds pressure for 20 minutes. Higher temperatures cause malt sugars to caramelize, preventing the agar from jelling properly.




http://www.shroomery.org/8506/Culture-Mediums


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OfflineTEAMSESH
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Re: germination problem [Re: Appalachian Brony]
    #22055471 - 08/06/15 08:38 PM (8 years, 5 months ago)

didn't sterilize the plates.  only the mea in a mason jar got serialized at some 15-16 psi


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OfflinePsilosoulful

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Re: germination problem [Re: Mad Season]
    #22055479 - 08/06/15 08:40 PM (8 years, 5 months ago)

I'm confused, are you saying that 10 psi is recommended for pasty plates then? Never made them before, so I don't know how they look like after a PC cycle. Just want to make sure I do it right the first time.


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OfflineTEAMSESH
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Re: germination problem [Re: Mad Season]
    #22055488 - 08/06/15 08:42 PM (8 years, 5 months ago)

Tanks ill give it a read :thumbup:


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OfflineTEAMSESH
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Re: germination problem [Re: TEAMSESH]
    #22055503 - 08/06/15 08:44 PM (8 years, 5 months ago)

great info hear I'm:thumbup: learning


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InvisibleMad Season
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Re: germination problem [Re: Psilosoulful]
    #22055514 - 08/06/15 08:47 PM (8 years, 5 months ago)

Quote:

Psilosoulful said:
I'm confused, are you saying that 10 psi is recommended for pasty plates then? Never made them before, so I don't know how they look like after a PC cycle. Just want to make sure I do it right the first time.



15 is fine. It's just  pulled from a lab book. It's just saying you don't want to overcook it. As bod said 15-20 minutes is fine. Going over 20 minutes, especially with malt agar would cause it to caramelize


--------------------
contam and car window art
How to shroomery like a pro! (Seriously, everyone read this!)
Improve your sterile techniques! (A comprehensive guide to agar)
Links upon links of literally EVERYTHING UP TO DATE

AMU Q&A
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OfflineTEAMSESH
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Re: germination problem [Re: bodhisatta]
    #22055566 - 08/06/15 08:57 PM (8 years, 5 months ago)

hey so the food grade agar im using is in a block forum.
weighs 10grams should I figure my own ratio to get it percept because don't know any thing that works with this agar .I've used powder in the past with 100% successes even brf agar style teks with successes. im using light malt extract to.


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OfflinePsilosoulful

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Re: germination problem [Re: Mad Season]
    #22055795 - 08/06/15 09:46 PM (8 years, 5 months ago)

Quote:

Mad Season said:
Quote:

Psilosoulful said:
I'm confused, are you saying that 10 psi is recommended for pasty plates then? Never made them before, so I don't know how they look like after a PC cycle. Just want to make sure I do it right the first time.



15 is fine. It's just  pulled from a lab book. It's just saying you don't want to overcook it. As bod said 15-20 minutes is fine. Going over 20 minutes, especially with malt agar would cause it to caramelize



Awesome, can't wait to mix up some agar!! :thumbup:


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OfflineTEAMSESH
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Re: germination problem [Re: Psilosoulful]
    #22056523 - 08/07/15 02:45 AM (8 years, 5 months ago)

If carmalization is to occur what effects would it have on germination?


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InvisibleMunchauzen
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Re: germination problem [Re: TEAMSESH]
    #22056536 - 08/07/15 02:53 AM (8 years, 5 months ago)

I PC my plates for 90 minutes. Never had any caramalization. Also never used malt dextrose extract. I do standard potato flakes or grain water recipes. they look the same before pc as they do after. you can pour them and let them cool down to look at them before you load them. mine usually have a nice glossy sheen to them.


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OfflinePsilosoulful

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Re: germination problem [Re: Munchauzen]
    #22057392 - 08/07/15 09:08 AM (8 years, 5 months ago)

Quote:

Munchauzen said:
I PC my plates for 90 minutes. Never had any caramalization.



:orry:


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InvisibleMad Season
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Re: germination problem [Re: Psilosoulful]
    #22057428 - 08/07/15 09:13 AM (8 years, 5 months ago)

Lol gw and flakes isn't malt. In the link I provided, malt was the only one mentioned that you had to worry about caramelization. If we used honey (which you totally can), it too can caramelize. I generally pc gw for 45, though I know 30 minutes is more than enough.


--------------------
contam and car window art
How to shroomery like a pro! (Seriously, everyone read this!)
Improve your sterile techniques! (A comprehensive guide to agar)
Links upon links of literally EVERYTHING UP TO DATE

AMU Q&A
No trees were killed in the sending of this message. However, a large number of electrons were terribly inconvenienced.


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