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PetMonkey



Registered: 05/24/20
Posts: 199
Last seen: 8 months, 13 days
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Help me understand Agar a little bit better
#26774581 - 06/22/20 03:45 PM (3 years, 7 months ago) |
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Hello, beautiful people.
My pet monkey is currently doing a PF/BRF cake grow (his second grow, first one was a few years ago and was a one-time, not-so-serious thing), but is thinking of moving onto agar/grain for the long term. Been reading a bunch of TEKs here and watching the video series from the legendary Roger Rabbit, but had a couple little things he wanted clarification on. Tell me if this is all correct, if you would:
- You can mix up enough agar to pour 10 plates, then clone a fresh mushroom or drop solution from a spore syringe into the plates
- Once the mycelia begin to colonize, you can split them in half and move them onto another 10 plates, giving you 20 plates total
- You can cut a wedge from one of your plates to colonize a master grain jar
- You can then use a single master grain jar to colonize 10 additional jars of grain
- You still have 20 plates with colonized mycelia sitting in a lunchbox in your fridge for when you want to do it all over again
Are all these statements correct? Sounds almost too good to be true, so I just wanted to make sure...
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Jus
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Re: Help me understand Agar a little bit better [Re: PetMonkey]
#26774643 - 06/22/20 04:09 PM (3 years, 7 months ago) |
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Quote:
You can mix up enough agar to pour 10 plates, then clone a fresh mushroom or drop solution from a spore syringe into the plates
- Once the mycelia begin to colonize, you can split them in half and move them onto another 10 plates, giving you 20 plates total
If you have perfectly clean samples or syringes then sure, this could work, but the reality is that you're probably going to get contamination, especially at first as you learn to nail the technique. I'm not there yet after a few batches.
When you transfer, you can select a section free from contamination (posting here can help select sections, it helps to have pictures of the plates lit well on a dark background and then held up to the light). A tiny section can be cut, some people use a hole punch or some other tool to do it, some just cut with a scalpel.
You can keep making strategic transfers to isolate or you can just use mixed genetics to spawn to bulk, which is fine, you just won't get those even grows that can consistently make huge fruiting bodies.
Everything else sounds fine to me, but I'm just starting out in the hobby so wait for another expert to confirm.
Edited by Jus (06/22/20 04:09 PM)
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EntheoGod
Entheo



Registered: 03/06/15
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Re: Help me understand Agar a little bit better [Re: Jus]
#26774651 - 06/22/20 04:14 PM (3 years, 7 months ago) |
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Just because a plate contams doesn’t mean it’s lost😊. You can transfer clean mycelium from agar and place it on a new plate to get healthy growth. Ever since starting agar myself I will never discourage anyone from doing it. It’s very forgiving and simple.
Agar is FOR helping you get clean spawn. Gotta start with something like either a spore print or spore syringe to get anywhere and by placing it on agar once you follow the right tek, you can clean up your act super fast. (Relative to how long brf takes or wbs straight from ms syringe.)
I’d recommend any beginner start with agar and learn that. It will help them develop there sterile techniques and practice makes perfection.
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PetMonkey



Registered: 05/24/20
Posts: 199
Last seen: 8 months, 13 days
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Re: Help me understand Agar a little bit better [Re: Jus]
#26775758 - 06/23/20 01:28 AM (3 years, 7 months ago) |
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Quote:
Jus said:
Quote:
You can mix up enough agar to pour 10 plates, then clone a fresh mushroom or drop solution from a spore syringe into the plates
- Once the mycelia begin to colonize, you can split them in half and move them onto another 10 plates, giving you 20 plates total
If you have perfectly clean samples or syringes then sure, this could work, but the reality is that you're probably going to get contamination, especially at first as you learn to nail the technique. I'm not there yet after a few batches.
When you transfer, you can select a section free from contamination (posting here can help select sections, it helps to have pictures of the plates lit well on a dark background and then held up to the light). A tiny section can be cut, some people use a hole punch or some other tool to do it, some just cut with a scalpel.
You can keep making strategic transfers to isolate or you can just use mixed genetics to spawn to bulk, which is fine, you just won't get those even grows that can consistently make huge fruiting bodies.
Everything else sounds fine to me, but I'm just starting out in the hobby so wait for another expert to confirm.
Understood, thanks for your input.
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PetMonkey



Registered: 05/24/20
Posts: 199
Last seen: 8 months, 13 days
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Re: Help me understand Agar a little bit better [Re: EntheoGod]
#26775760 - 06/23/20 01:30 AM (3 years, 7 months ago) |
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Quote:
EntheoGod said: Just because a plate contams doesn’t mean it’s lost😊. You can transfer clean mycelium from agar and place it on a new plate to get healthy growth. Ever since starting agar myself I will never discourage anyone from doing it. It’s very forgiving and simple.
Just curious, what do you do with a petri dish that has been contaminated after the fact? You cut out a section of clean mycelium to transfer to a new sterile dish, but then how do you clean out/sterilize the old dish? I've read that polyurethane petri dishes can't handle a cycle in a pressure cooker, so just wondering how else you would do it? Thanks for your reply.
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Jus
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Re: Help me understand Agar a little bit better [Re: PetMonkey]
#26775831 - 06/23/20 03:10 AM (3 years, 7 months ago) |
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Quote:
PetMonkey said: Just curious, what do you do with a petri dish that has been contaminated after the fact? You cut out a section of clean mycelium to transfer to a new sterile dish, but then how do you clean out/sterilize the old dish? I've read that polyurethane petri dishes can't handle a cycle in a pressure cooker, so just wondering how else you would do it? Thanks for your reply. 
Those types of petri dishes are generally disposable. You can get glass dishes that are more expensive. I've got some that can only handle 160C, so need to be dry heated (although there's water present from the washing I do so it's actually steam heating initially) for over 2 hours.
You might be able to get other dishes which can be pressure cooked, shortening the time.
You might want to look into no pour techniques where you can use small cheapish tubs, pre-filled with agar, and pressure cook.
I clean my contaminated glass dished by scraping it into a bin and then washing them in hot soapy water, scrubbing well.
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EntheoGod
Entheo



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Re: Help me understand Agar a little bit better [Re: Jus]
#26775883 - 06/23/20 04:56 AM (3 years, 7 months ago) |
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PetMonkey



Registered: 05/24/20
Posts: 199
Last seen: 8 months, 13 days
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Re: Help me understand Agar a little bit better [Re: EntheoGod]
#26777168 - 06/23/20 04:03 PM (3 years, 7 months ago) |
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Quote:
EntheoGod said:
Plus if theyre contaminated and youre done with the plate, clean it out with soap and hot water then bam youre good but you could also use a bleach water solution.
[
I know you said you don't typically use petri dishes, but after washing with soap and hot water, how do you store them safely? Couldn't they hypothetically get contaminates as they're sitting on a towel drying out?
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mushpunx
Fungus Punk


Registered: 04/20/14
Posts: 13,394
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Re: Help me understand Agar a little bit better [Re: PetMonkey]
#26777207 - 06/23/20 04:17 PM (3 years, 7 months ago) |
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Spore prints are taken from fruits that have been grown in open air, therefore they are not going to be 100% sterile. By extension , spore syringes made from those prints will never be 100% sterile either. Syringes made from prints taken from indoor fruits are usually clean enough to put to BRF cakes, if used for grains you will run into contamination problems sooner or later.
Agar gives us a tool to isolate clean mycelium away from contaminants carried along with the spores. It gives us the ability to grow clean innoculant.
In addition to giving us a 2-D surface to separate pure mycelium from contaminants, agar also let's us expand and store our live cultures, isolate strains and clone.
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 Amateur Mycologists United AMU Q&A
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EntheoGod
Entheo



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Re: Help me understand Agar a little bit better [Re: PetMonkey]
#26777259 - 06/23/20 04:35 PM (3 years, 7 months ago) |
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Youre going to be sterilizing them in a pressure cooker. I dont think it would matter where u store them as long as you clean them properly before hand.
PCing should kill anything that would be on them as far as I know. Again, I have made one batch of agar and so far so good. I didnt worry about it after I cleaned em with soap and water just be sure you dont get any large debris inside them before you pour the agar inside.
Once you make the agar and place it in the containers youre going to pressure cook them for the amount of time called for in the tek, I cant remember off the top of my head atm.
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Jus
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Re: Help me understand Agar a little bit better [Re: PetMonkey]
#26778071 - 06/23/20 11:00 PM (3 years, 7 months ago) |
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Quote:
PetMonkey said:
Quote:
EntheoGod said:
Plus if theyre contaminated and youre done with the plate, clean it out with soap and hot water then bam youre good but you could also use a bleach water solution.
[
I know you said you don't typically use petri dishes, but after washing with soap and hot water, how do you store them safely? Couldn't they hypothetically get contaminates as they're sitting on a towel drying out?
My general workflow for cleaning and sterilising is to wash them well in hot soapy water, shake off excess water, then transfer immediately onto a double layer of foil. I stack the plates 5 high (that's how big my SAB is), wrap them tightly with the two layers of foil, and then cook at 160C for over 2 hours.
After they've cooled down, they should be sterile in the foil, which can be placed into a SAB and carefully unwrapped, letting air settle afterwards.
So far I've only gotten a single contamination from the plates, and that was when I was trying to dry them with paper towels before wrapping in foil, a pointless exercise that only served to make the plates less clean before going in and increased the sterilisation time by removing all the water so it couldn't evaporate into steam in the oven.
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mushpunx
Fungus Punk


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Re: Help me understand Agar a little bit better [Re: Jus]
#26778327 - 06/24/20 02:42 AM (3 years, 7 months ago) |
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You are using glass resusable plates, yeah?
I've heard of people oven sterilizing them. Why not just PC though?
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 Amateur Mycologists United AMU Q&A
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Jus
Stranger


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Re: Help me understand Agar a little bit better [Re: mushpunx]
#26778713 - 06/24/20 08:34 AM (3 years, 7 months ago) |
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Quote:
mushpunx said: You are using glass resusable plates, yeah?
I've heard of people oven sterilizing them. Why not just PC though?
Honestly, because I misremembered the temperature/pressure graphs and way overestimated the temperature inside a pressure cooker.
I think I'll switch to pressure cooking...
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PetMonkey



Registered: 05/24/20
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Last seen: 8 months, 13 days
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Re: Help me understand Agar a little bit better [Re: mushpunx] 1
#26782455 - 06/25/20 04:09 PM (3 years, 7 months ago) |
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Quote:
mushpunx said: You are using glass resusable plates, yeah?
I've heard of people oven sterilizing them. Why not just PC though?
Nice avatar Mush, looks like Golden Teachers?
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Nephedryn
Apothecary



Registered: 11/29/18
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Re: Help me understand Agar a little bit better [Re: PetMonkey] 1
#26782481 - 06/25/20 04:17 PM (3 years, 7 months ago) |
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Use this tek and skip the g2g transfers, next level tek right here by MudaFucka. https://www.shroomery.org/forums/showflat.php/Number/20429745
-------------------- The point singularity of the shroomery
Ultimate Tek Compendium: For Noobs And Onward “One glance at a book and you hear the voice of another person, perhaps someone dead for 1,000 years. To read is to voyage through time.”
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mushpunx
Fungus Punk


Registered: 04/20/14
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Re: Help me understand Agar a little bit better [Re: PetMonkey] 1
#26783328 - 06/25/20 09:53 PM (3 years, 7 months ago) |
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Quote:
PetMonkey said:
Quote:
mushpunx said: You are using glass resusable plates, yeah?
I've heard of people oven sterilizing them. Why not just PC though?
Nice avatar Mush, looks like Golden Teachers?
Thanks! That was a few years ago, it was AA Albinos, or A+ , I can't remember
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 Amateur Mycologists United AMU Q&A
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mushpunx
Fungus Punk


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Re: Help me understand Agar a little bit better [Re: Nephedryn]
#26783334 - 06/25/20 09:58 PM (3 years, 7 months ago) |
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Quote:
Nephedryn said: Use this tek and skip the g2g transfers, next level tek right here by MudaFucka. https://www.shroomery.org/forums/showflat.php/Number/20429745
I made LI blenders before with the Oster parts. If you search eBay eventually you will find used Eberbach units for sale (They are fully autoclavable and meant for this purpose). I have two, a stainless steel 360ml one (they're about $600 new), and a glass 500ml one, ($300 new). I got each for less than $30. Same with the Waring lab blender + seconds timer

It'll take a minute but you'll find them being sold cheap eventually
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 Amateur Mycologists United AMU Q&A
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