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RonPaulVerm
Piss Christ


Registered: 07/21/08
Posts: 1,072
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tissue doesn't like peroxide agar?
#11697364 - 12/22/09 06:51 AM (14 years, 1 month ago) |
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So I made some PDYA dishes and while the agar was cooling in the bottles I put in a little less than a cap of H2O2 in each bottle. (they were half-filled Arizona Ice Tea bottles) I figured it would be fine because I figure 6-10 ml per liter of agar liquid.
My transferred tissues are doing great. They're getting nice & fuzzy and sending out those fine hairs to latch onto the agar. It's been about 5 days for some of the plates. However, they aren't attaching to the plate that fast. Could the peroxide be affecting their affection for the agar?
Other details: I have my unused plates stored in a fridge and take them out and let them get to room temp before using I used a drop of food coloring to make the agar mixture all pink
I figured I could just wait it out, but a thread about this might help someone in the future
-------------------- Spawn I Have: Blewitt, Reishi, Maitake, Chicken of the Woods, Parasol, Shaggy Mane, King, Blue and Gold Oyster, Shiitake, King Stropharia, Lions Mane, Almond Portabello, Elm Oyster, Phoenix Oyster, Nameko, Enoki, White Beech, Spawn I Want: Corcyceps Sp., Cauliflower Mushrooms, Agaricus species (Portobello), Pink Oyster, Piopinno, ...anything else you might have Primitive Skills Any information I've posted is only related to fictional purposes. I do not advocate growing any illegal mushrooms ...I DO advocate learning about Law Enforcement Against Prohibition
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Ogla



Registered: 02/16/04
Posts: 11,314
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Re: tissue doesn't like peroxide agar? [Re: RonPaulVerm]
#11697494 - 12/22/09 08:01 AM (14 years, 1 month ago) |
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I dont think its necessary to add peroxide to the agar mix. I believe its meant to sterilize the surface of a shroom when ur cloning tissue to agar.
if u nuked it in the microwave, that should be enough to have sterlize it by itself.
Based on what ur saying, the peroxide may have killed some of your tissues and that may be why ur not seeing as aggressive growth on some on those plates. However, the peroxide should turn into water eventually.
I dont have much experince on dishes yet, but am currently working on some agar plates anyways. Good luck
Edited by Ogla (12/22/09 08:06 AM)
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Cloneufc
Master Exploder!



Registered: 11/15/07
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Loc: Las Vegas
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Re: tissue doesn't like peroxide agar? [Re: Ogla]
#11697585 - 12/22/09 08:29 AM (14 years, 1 month ago) |
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Peroxide should only be used for killing cobweb mold. I have not used peroxide for anything else in my almost 4 years of growing. It can kill the tissue and is not needed.
Edited by Cloneufc (12/22/09 08:30 AM)
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RogerRabbit
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Registered: 03/26/03
Posts: 42,214
Loc: Seattle
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Re: tissue doesn't like peroxide agar? [Re: Cloneufc]
#11697626 - 12/22/09 08:42 AM (14 years, 1 month ago) |
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Peroxide is toxic to fungi. Don't use it in agar, as it will impede growth. I know it's been recommended around here by others, but not by me. RR
-------------------- Download Let's Grow Mushrooms semper in excretia sumus solim profundum variat "I've never had a failed experiment. I've only discovered 10,000 methods which do not work." Thomas Edison
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RonPaulVerm
Piss Christ


Registered: 07/21/08
Posts: 1,072
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Re: tissue doesn't like peroxide agar? [Re: RogerRabbit]
#11697646 - 12/22/09 08:50 AM (14 years, 1 month ago) |
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I hear ya, but even Paul Stamets talks about peroxide treating substrate so that the mycelium will get a head start to contaminants...there is a whole section in Mycelium Running. Not that he is the know all end all but I guess he's experienced success with it. In this thread both Roadkill and Scatmanrav talk about adding peroxide to agar. Scatmanrav mentions that i will break down and Roadkill is quoting R. Rush Wayne Ph.D.
And now that I read it Roadkill syas that the more peroxide the slower the growth. So I guess I should just be patient.
-------------------- Spawn I Have: Blewitt, Reishi, Maitake, Chicken of the Woods, Parasol, Shaggy Mane, King, Blue and Gold Oyster, Shiitake, King Stropharia, Lions Mane, Almond Portabello, Elm Oyster, Phoenix Oyster, Nameko, Enoki, White Beech, Spawn I Want: Corcyceps Sp., Cauliflower Mushrooms, Agaricus species (Portobello), Pink Oyster, Piopinno, ...anything else you might have Primitive Skills Any information I've posted is only related to fictional purposes. I do not advocate growing any illegal mushrooms ...I DO advocate learning about Law Enforcement Against Prohibition
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RogerRabbit
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Registered: 03/26/03
Posts: 42,214
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Re: tissue doesn't like peroxide agar? [Re: RonPaulVerm]
#11697667 - 12/22/09 09:01 AM (14 years, 1 month ago) |
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The whole idea of using petri dishes is to encourage rapid growth of mycelium in order to isolate it away from contaminants or other strains. Using peroxide in any amount degrades the growth of the culture, defeating the purpose. It's better to make a series of transfers to isolate your culture, rather then to depend on peroxide, which is as toxic to your mycelium as it is to other species of mycelium or bacteria. Without the peroxide, you'd have already made two or three transfers by now and be finished. We have to be careful relying on 20 year old growing advice when we've advanced the hobby beyond that point. I'm sure your culture will begin to colonize the agar once the peroxide breaks down, but then what's the point of having used it in the first place? RR
-------------------- Download Let's Grow Mushrooms semper in excretia sumus solim profundum variat "I've never had a failed experiment. I've only discovered 10,000 methods which do not work." Thomas Edison
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RonPaulVerm
Piss Christ


Registered: 07/21/08
Posts: 1,072
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Re: tissue doesn't like peroxide agar? [Re: RogerRabbit]
#11697740 - 12/22/09 09:23 AM (14 years, 1 month ago) |
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I would agree RR. Well I have over 30 plates left that I think I will use anyway and make master slants out of becuse like you said they aren't ideal for transfers. I don't think I will use peroxide again though this has been a learning experience. It gave me hopes though because last batch of plates I poured (though i was sloppy beyond my standards) had almost half contam rate AND THEY HADN"T BEEN USED haha. Well thanks for the input and you can bet I won't use peroxide in agar again because it's using excess materials, causing more waiting, and harmful.
TWO questions though: -The plates I haven't used and might not for use up for a couple of weeks...you think the peroxide will break down while they're in the fridge? -Does the peroxide just slow growth or does it also diminish the quality of the species?
-------------------- Spawn I Have: Blewitt, Reishi, Maitake, Chicken of the Woods, Parasol, Shaggy Mane, King, Blue and Gold Oyster, Shiitake, King Stropharia, Lions Mane, Almond Portabello, Elm Oyster, Phoenix Oyster, Nameko, Enoki, White Beech, Spawn I Want: Corcyceps Sp., Cauliflower Mushrooms, Agaricus species (Portobello), Pink Oyster, Piopinno, ...anything else you might have Primitive Skills Any information I've posted is only related to fictional purposes. I do not advocate growing any illegal mushrooms ...I DO advocate learning about Law Enforcement Against Prohibition
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RogerRabbit
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Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 11 months, 2 days
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Re: tissue doesn't like peroxide agar? [Re: RonPaulVerm]
#11697817 - 12/22/09 09:44 AM (14 years, 1 month ago) |
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I wouldn't use those plates that the mycelium doesn't want to grow on. I also generally toss out unused plates after two weeks or so. Trying to use them beyond that point is pushing luck. RR
-------------------- Download Let's Grow Mushrooms semper in excretia sumus solim profundum variat "I've never had a failed experiment. I've only discovered 10,000 methods which do not work." Thomas Edison
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