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jaydeezy817
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Agar dish airflow
#26990331 - 10/17/20 04:37 PM (3 years, 3 months ago) |
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Im prepping condiment cups for agar. They are air tight. Should i add airflow to these for mycelium? Probably a dumb question. Thanks
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Rhizomorph
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Yeah they should have some form of gas exchange. Just a hole with 2-3 layers of some micropore tape works.
See the section "modifying the containers" in this write-up by Josex: https://www.shroomery.org/forums/showflat.php/Number/25137693
I also highly recommend the holy grail plates in my signature since they require no modification but that's up to you.
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jaydeezy817
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Thanks. Very much appreciated
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Rhizomorph
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For sure - Good luck!!
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jaydeezy817
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Im so bad at agar. Is this contamination or what.
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FoTwenty
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Contaminated.....
-------------------- "I have realized that the past and future are real illusions, that they exist in the present, which is what there is and all there is.” ― Alan Wilson Watts

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Rhizomorph
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yup, contaminated. How long did you PC them for?
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jaydeezy817
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Fuck. Only 30 minutes at 15 psi. Agar keeps kicking my ass for years now. Should i completely sterilize for 2 hours?
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Rhizomorph
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30min at 15 psi should be more than enough but if you want to be extra safe you could go up to 40min. Anything beyond that risks caramelizing the agar.
Did you open the containers after pulling them out of the PC? If you inoculated them with a syringe for example it could have come from the syringe. Otherwise there should be no reason one of those containers should get contamination if you PC'd for 30min @15 psi and haven't opened the containers since.
Another thing you could try is letting the tape dry off before pulling them out. Although contaminants get caught in the tape when air passes through, if the tape is wet it allows contaminants to crawl through the micropore. I've never had an issue with it if you allow for proper drying but it is possible. Putting the micropore on the side instead of the tops of the containers can help too as contaminants generally fall downwards rather than sideways.
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Brian Jones
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30 minutes at 15psi is fine and that didn't cause the problem.
-------------------- "The Rolling Stones will break up over Brian Jones' dead body" John Lennon I don't want no commies in my car. No Christians either. The worst thing about corruption is that it works so well,
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jaydeezy817
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Man thanks guys. You really help me with this info..I just cant think where the contam comes from because i use the same techniques i use with my grain and never have contam issues with those.... I think im gonna get shallow glass jars instead of ketchup cups and use smaller needles with syringes because even after flame sterilizing the solution will drip out and leave the needle wet..also im gonna gonna use synthetic filter discs instead of tape..i will post update if anyone wants to check back here but any info anyone has is greatly appreciated.
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jaydeezy817
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Quote:
jaydeezy817 said: Man thanks guys. You really help me with this info..I just cant think where the contam comes from because i use the same techniques i use with my grain and never have contam issues with those.... I think im gonna get shallow glass jars instead of ketchup cups and use smaller needles with syringes because even after flame sterilizing the solution will drip out and leave the needle wet..also im gonna gonna use synthetic filter discs instead of tape..i will post update if anyone wants to check back here but any info anyone has is greatly appreciated.
Instead of glass jars thinking glad mini rounds or any small container and just sterilize in dishes
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jaydeezy817
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Your holy grail plates are genius
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Rhizomorph
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They're not mine. I just swear by them
Mr. Alien is the brilliant person who started using them for agar.
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Rafiikii



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if you say inoculated grain comes out clean, then work on your sterile technique granted your syringes are clean.
Do you have a still air box?
the cool thing about agar is you work away from the contamination, as long as you have some mycelial germination of course.
-------------------- "You didn’t come into this world. You came out of it, like a wave from the ocean. You are no stranger here."  
Edited by Rafiikii (10/23/20 09:32 PM)
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TheBoJim
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I wrap my dishes with cling wrap, so no GE I am assuming, and never have a problem. Is GE really necessary for what we do?
Quote:
Rhizomorph said: Yeah they should have some form of gas exchange. Just a hole with 2-3 layers of some micropore tape works.
See the section "modifying the containers" in this write-up by Josex: https://www.shroomery.org/forums/showflat.php/Number/25137693
I also highly recommend the holy grail plates in my signature since they require no modification but that's up to you. 
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Rhizomorph
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Re: Agar dish airflow [Re: TheBoJim]
#27000637 - 10/23/20 11:08 PM (3 years, 3 months ago) |
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Cling wrap is actually breathable (for mushrooms, not for humans) depending on the brand, so you're probably getting mild gas exchange - I assume it is Glad Cling Wrap? mild is the key word. It needs to breath, but probably very little.
There's probably enough air even in an airtight dish for a colony to grow, but it will grow better with a minimal amount of gas exchange versus none at all. Don't quote me on this though - I've never tried making a plate without any gas exchange.
Each container is going to be different too. petri dishes are not airtight so wrapping them in cling wrap allows gas exchange but the cling wrap prevents contaminants from getting in. Most ketchup cups do tend to be airtight (to my knowledge) and thus they require modification.
What I love about the HG plates I mentioned is that the lids themselves are breathable but because the screw-top design creates an obscure pathway, contaminants can't get in. So the lids themselves allow gas exchange while keeping out contaminants. No cling wrap or punching holes needed!
Edited by Rhizomorph (10/23/20 11:14 PM)
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gt40
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Quote:
jaydeezy817 said: Man thanks guys. You really help me with this info..I just cant think where the contam comes from because i use the same techniques i use with my grain and never have contam issues with those.... I think im gonna get shallow glass jars instead of ketchup cups and use smaller needles with syringes because even after flame sterilizing the solution will drip out and leave the needle wet..also im gonna gonna use synthetic filter discs instead of tape..i will post update if anyone wants to check back here but any info anyone has is greatly appreciated.
I read the info on the page of well known spore vendor that 99% times when spore - agar shows contams, same spore to grain will not and you can success. It sounds suprisingly because local growing culture insists that you must(!) use agar first. So if it works for you also just keep going
Edited by gt40 (10/24/20 12:16 AM)
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