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wolfXsoul
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Agar: What Am I Doing Wrong??
#26855814 - 07/31/20 04:44 PM (3 years, 6 months ago) |
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Background: I've been trying to get into agar, but keep getting a fairly high contam rate before even using the plates! Ive tried 2 different containers and after a few days they all look like this, before even opening them up. The only thing I can think is that my PC isn't getting to high enough temp? I use an instapot but have had no problems with my grain. Any advice/suggestions?

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Rhizomorph
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Re: Agar: What Am I Doing Wrong?? [Re: wolfXsoul]
#26855922 - 07/31/20 05:48 PM (3 years, 6 months ago) |
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Are you keeping the lids on after pressure cooking? The bacteria appears to be spreading from the outside to the inside. This makes me think that it is not from the air, but rather, it might be climbing down the sides of the containers - you could try saran wrap/parafilming/micropore taping along the edges of the lid and if this makes a difference you can pinpoint the source being from the edges of the lid.
Otherwise I would advise trying a different pressure cooker. I've heard conflicting info on instapots but from my understanding you should avoid them. I don't know about this for certain though. Read the manual or search online and find if it reaches 15psi.
Edited by Rhizomorph (07/31/20 05:51 PM)
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skullhuman
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Re: Agar: What Am I Doing Wrong?? [Re: Rhizomorph]
#26855926 - 07/31/20 05:50 PM (3 years, 6 months ago) |
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Must not be sterilizing properly.
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TheDoobsker
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Re: Agar: What Am I Doing Wrong?? [Re: wolfXsoul]
#26855928 - 07/31/20 05:51 PM (3 years, 6 months ago) |
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An instapot should sterilize sufficiently. That's what I use for agar and have never had issues that seemed to be caused by it. It's possible there's an issue with yours, but that's easy to check; see if it cooks a meal for you without issues. It should cook any type of food in about half the time that a stovetop would.
Correct me if I'm wrong, but I don't think those containers will survive a PC, so I assume you're PC'ing the agar and then pouring into the containers?
If so, how are you sterilizing the containers and what is your method of pouring?
This is probably where your bacteria is entering.
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Rhizomorph
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Re: Agar: What Am I Doing Wrong?? [Re: TheDoobsker]
#26855932 - 07/31/20 05:53 PM (3 years, 6 months ago) |
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PP5 plastic can survive pressure cooking (look at the little recycling logo in the pictures with "PP" below it and a "5" in the middle indicates it is PP5)
Most no-pour methods use some variation of PP5 plastic containers.
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Gan
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Re: Agar: What Am I Doing Wrong?? [Re: Rhizomorph]
#26855962 - 07/31/20 06:11 PM (3 years, 6 months ago) |
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How long are you sterilizing in your instapot for?
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TheDoobsker
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Re: Agar: What Am I Doing Wrong?? [Re: Rhizomorph]
#26855978 - 07/31/20 06:19 PM (3 years, 6 months ago) |
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Quote:
Rhizomorph said: PP5 plastic can survive pressure cooking (look at the little recycling logo in the pictures with "PP" below it and a "5" in the middle indicates it is PP5)
Most no-pour methods use some variation of PP5 plastic containers.
Aah yeah you're right. I've learned that pp5 can be PC'd but didn't look at the logo. I just saw cheap plastic ketchup bottles and assumed they must have been poured. Thanks for the correction 
As far as the instapot discussion, I don't think that's a problem if it's working correctly. I leave 1-2 plates un-inoculated with every batch of agar I make and let them sit as a test. I've never seen a contam in those plates. It's definitely sterilizing just fine.
However, instapots have a working pressure of 10.2-11.6 psi which obviously is concerning. But like I said, they work.
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sandman420
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Re: Agar: What Am I Doing Wrong?? [Re: TheDoobsker]
#26856084 - 07/31/20 07:32 PM (3 years, 6 months ago) |
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If agar looks like that in 2 days than the answer is almost definitely that you didnt actually sterilize the agar. Even drawing air in while cooling will take a week to look like that and bacteria isnt really that airborne you would see a lot of mold too.
In agar, bacteria is typically spread from touching something dirty or coming off the grower. But in this case its looking like just not sterilized.
You are probably not venting the PC.
Edited by sandman420 (07/31/20 07:36 PM)
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sandman420
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Re: Agar: What Am I Doing Wrong?? [Re: sandman420]
#26856090 - 07/31/20 07:35 PM (3 years, 6 months ago) |
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oh its an instapot go figure yea not sterilizing sounds like.
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TheDoobsker
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Re: Agar: What Am I Doing Wrong?? [Re: sandman420]
#26856248 - 07/31/20 08:55 PM (3 years, 6 months ago) |
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Quote:
sandman420 said: oh its an instapot go figure yea not sterilizing sounds like.
I don't mean to sound rude, but have you ever actually used one or seen real evidence that they don't work? Let's not forget that everything we've learned about science is a result of experiments.
They're less efficient than a real PC based on the pressure difference. But do you really think that much bacteria would survive 210F & 11psi for 30 minutes? I think it's more of an assurance thing. These conditions will likely kill everything whereas at 15psi, it's not even a question. Nothing will live.
I doubt these containers sat through 30 mins of 210F & 11psi and grew that much bacteria within 3 days. There's probably another factor going on here.
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sandman420
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Re: Agar: What Am I Doing Wrong?? [Re: TheDoobsker]
#26856277 - 07/31/20 09:17 PM (3 years, 6 months ago) |
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Im just saying its not a pressure cooker that is manually operated so who know exactly what is going on in there
anyone can look at that agar and say it is almost definitely contammed because it wasnt sterilized. Such a short time no way it could be anything else.
Im not saying an instapot couldnt work.
I am saying his probably isnt working good enough, is the most likely problem here. Maybe sensor malfunction? Maybe cheap chinese piece of shit that has a 10% malfunction rate? Who knows.
Edited by sandman420 (07/31/20 09:25 PM)
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sandman420
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Re: Agar: What Am I Doing Wrong?? [Re: TheDoobsker]
#26856290 - 07/31/20 09:26 PM (3 years, 6 months ago) |
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Quote:
TheDoobsker said: [
I doubt these containers sat through 30 mins of 210F & 11psi and grew that much bacteria within 3 days.
Bingo they didnt.
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Enkidu
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Re: Agar: What Am I Doing Wrong?? [Re: sandman420]
#26856326 - 07/31/20 09:48 PM (3 years, 6 months ago) |
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Prep just the agar and pc it then pour plates in an sab
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c10h12n2o
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Re: Agar: What Am I Doing Wrong?? [Re: Enkidu]
#26856573 - 08/01/20 01:59 AM (3 years, 6 months ago) |
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Instapot can work but you probably need to run it much longer than a pc, because of thr lower pressure
But i wouldnt use one. I usually do agar at 17-20 psi for 20 mins
Make some control plates to figure out where you are going wrong. Might be improper sterilization, wrong containers, unsterile inoculation, improper handling, etc. If the control plates dont contam it means sterilization worked
--------------------
  C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide "Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing." "Convictions are more dangerous enemies of truth than lies" ― Friedrich Nietzsche
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pesa



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Re: Agar: What Am I Doing Wrong?? [Re: wolfXsoul]
#26856575 - 08/01/20 02:02 AM (3 years, 6 months ago) |
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1. cut the nutrients in half if you are using malt at 10 g bring it down to 5g. more nutrients means more food for bacteria. in cloning or spore germination agar does not have to be that much rich in nutrients.2:1 agar:nutrients
2. add a antibiotic like the one they sell for fish aquarium.
3. pressure cook for 20 to 25 minutes.
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c10h12n2o
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Re: Agar: What Am I Doing Wrong?? [Re: pesa]
#26856606 - 08/01/20 03:30 AM (3 years, 6 months ago) |
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The amount of nutrients in the agar isnt the problem.
He does not beed an antibiotic. If he is getting contamination before using the plates clearly there is an issue with sterilization and clean technique. There are very, very few cases where you need antibiotics, this isnt one of them
He doesn't have a legit pc, he has an instapot which is like 11psi. He needs to go much much longer than the 20 minutes it takes at 15-20psi
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  C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide "Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing." "Convictions are more dangerous enemies of truth than lies" ― Friedrich Nietzsche
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wolfXsoul
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Re: Agar: What Am I Doing Wrong?? [Re: c10h12n2o]
#26856676 - 08/01/20 06:07 AM (3 years, 6 months ago) |
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Thanks for all of the replies guys. A quick rundown: they are pp5 containers/lids. I've tried 2 different containers, so I dont think its the containers themselves. I've tried going as long as 2 hours on the instapot, and stil contams. I guess its simply not getting hot enough? How are my grain jars working just fine then? Maybe because they are enclosed in glass and getting hotter?
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sandman420
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Re: Agar: What Am I Doing Wrong?? [Re: wolfXsoul]
#26856761 - 08/01/20 07:20 AM (3 years, 6 months ago) |
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That is interesting information that you have been sterilizing grain no problem. Are you suuuuuuuure you've been sterilizing grain no problem as in successfully spawned some of this grain even 1 time?
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wolfXsoul
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Re: Agar: What Am I Doing Wrong?? [Re: sandman420]
#26856789 - 08/01/20 07:45 AM (3 years, 6 months ago) |
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I've spawned and fruited 24 qts with no contams as of yet. That's a win at least!
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sandman420
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Re: Agar: What Am I Doing Wrong?? [Re: wolfXsoul]
#26856805 - 08/01/20 08:01 AM (3 years, 6 months ago) |
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No doubt. hmm.
Well it is still possible that the unit is malfunctioning now. Has this been happening at the same time that the grain is successful?
Or has it been a minute since you did a good grain run?
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