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jacc1234
newbie
Registered: 09/30/02
Posts: 74
Last seen: 17 years, 2 months
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Using H2O2 to sterilize tissue before cloning.....
#5599236 - 05/06/06 10:32 PM (17 years, 8 months ago) |
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I think i remember reading in one of the Stament's books about people dipping a slice of the mushroom tissue in H2O2 before placing it on agar to help kill any possible surface contams. Yet, I have not seen any mention of this in the agar teks. Am I completly off or is this a viable way to limit contam risk even more.
Thanks, jacc1234
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shirley knott
not my real name

Registered: 11/11/02
Posts: 9,105
Loc: London
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Re: Using H2O2 to sterilize tissue before cloning..... [Re: jacc1234]
#5600056 - 05/07/06 02:57 AM (17 years, 8 months ago) |
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many growers use peroxidated agar, but still, a quick plunge in peroxide will be a good idea if contams are suspected. when i receive a wedge in the post, for example, i usually dunk it for 10 seconds before plonking it on peroxide agar.
-------------------- buh
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jacc1234
newbie
Registered: 09/30/02
Posts: 74
Last seen: 17 years, 2 months
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Re: Using H2O2 to sterilize tissue before cloning..... [Re: shirley knott]
#5600434 - 05/07/06 09:00 AM (17 years, 8 months ago) |
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Thanks, just needed to hear it from someone else.
Jacc1234
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Roker
Stranger



Registered: 11/02/05
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Loc: outer spiral arm
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Re: Using H2O2 to sterilize tissue before cloning..... [Re: shirley knott]
#5616318 - 05/11/06 05:54 AM (17 years, 8 months ago) |
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what volume of peroxide do you use for dunking shirley?
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hyphae
born to grow


Registered: 12/13/02
Posts: 6,228
Loc: the rain forests
Last seen: 12 years, 8 months
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Re: Using H2O2 to sterilize tissue before cloning..... [Re: Roker]
#5627960 - 05/14/06 10:37 AM (17 years, 8 months ago) |
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I dunk in straight 3%, I actually cut out a center section from the larger specimen under the peroxide when cloning.
-------------------- Getting the most out of your casings!, A pinning strategy. Oyster Shell "Flour" $2 for 1lb. a hell of a deal Not what is overlay but rather what overlay is Gas Exchange vs. FAE "We all have priorities. I used a closet once setup a nice little lab trouble was all the shit that was in there ended up in the bedroom that pissed off the GF then I ended up dumping her as she was getting in the way of my sterile culture technique! Ya I got priorities too!!!"
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ohmatic
searcher


Registered: 02/28/04
Posts: 6,742
Loc: europe
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Re: Using H2O2 to sterilize tissue before cloning..... [Re: hyphae]
#5628508 - 05/14/06 02:01 PM (17 years, 8 months ago) |
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under the peroxide ?
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MONOTUB tek HEATBOMB tek RIP #cultivation! ....can't associate? well FUCK U !
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hyphae
born to grow


Registered: 12/13/02
Posts: 6,228
Loc: the rain forests
Last seen: 12 years, 8 months
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Re: Using H2O2 to sterilize tissue before cloning..... [Re: ohmatic]
#5628856 - 05/14/06 03:44 PM (17 years, 8 months ago) |
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Submerged.
-------------------- Getting the most out of your casings!, A pinning strategy. Oyster Shell "Flour" $2 for 1lb. a hell of a deal Not what is overlay but rather what overlay is Gas Exchange vs. FAE "We all have priorities. I used a closet once setup a nice little lab trouble was all the shit that was in there ended up in the bedroom that pissed off the GF then I ended up dumping her as she was getting in the way of my sterile culture technique! Ya I got priorities too!!!"
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RogerRabbit
Bans for Pleasure


Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 11 months, 2 days
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Re: Using H2O2 to sterilize tissue before cloning..... [Re: hyphae]
#5630152 - 05/14/06 09:08 PM (17 years, 8 months ago) |
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If you still have problems with contaminants after using peroxide, you can also dip tissue into a ten percent bleach solution for a couple of minutes. Believe it or not, the mycelium can withstand this, but molds and bacteria can't. The bleach works really well when cloning dry tissue. Live tissue needs a week or two to recover from the bleach, but dry tissue is dormant so hardly even gets smacked. 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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ohmatic
searcher


Registered: 02/28/04
Posts: 6,742
Loc: europe
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Re: Using H2O2 to sterilize tissue before cloning..... [Re: RogerRabbit]
#5630626 - 05/14/06 11:18 PM (17 years, 8 months ago) |
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actually i gotta try playnig around with some dry tissue, just for kicks - the thought of mushies being able to still get back to living is just so bloody amazing
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MONOTUB tek HEATBOMB tek RIP #cultivation! ....can't associate? well FUCK U !
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waixingren


Registered: 03/14/05
Posts: 2,644
Loc: SW Florida
Last seen: 8 days, 2 hours
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Re: Using H2O2 to sterilize tissue before cloning..... [Re: RogerRabbit]
#5634181 - 05/15/06 06:34 PM (17 years, 8 months ago) |
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using dry tissue is definetly interesting. RR long do you suppose tissue could be dry before being beyond recovery?
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RogerRabbit
Bans for Pleasure


Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 11 months, 2 days
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Re: Using H2O2 to sterilize tissue before cloning..... [Re: waixingren]
#5634859 - 05/15/06 08:36 PM (17 years, 8 months ago) |
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I've recovered three year old tissue. There will definitely be contamination, but you'll be able to work through it with agar. I generally try to do transfers as soon as I see mycelium without waiting to see if it contaminates or not. At other times, bacteria or mold will start to grow first, then the mushroom mycelium. In this case, all you can do is begin a series of transfers until you have a pure culture. 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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Mushroplex
Mycology Enthusiast



Registered: 10/30/22
Posts: 35
Last seen: 9 months, 1 hour
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Re: Using H2O2 to sterilize tissue before cloning..... [Re: shirley knott]
#28110024 - 12/23/22 02:34 PM (1 year, 1 month ago) |
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Seems interesting has anyone been successful in cloning using this method ? BLEACH OR h202?
Edited by Mushroplex (12/23/22 02:36 PM)
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ohkw
Stranger

Registered: 07/12/22
Posts: 96
Loc: Austria
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Re: Using H2O2 to sterilize tissue before cloning..... [Re: Mushroplex]
#28110608 - 12/24/22 01:20 AM (1 year, 1 month ago) |
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Using H2O2 never worked for me. Depending on th etime of exposure, either the tissue sample was dead as well, or the contaminations grew even better ... I used pretty small samples. Maybe using pieces with more beef on it can work better. But if your sample is very old, it's likely that bacteria already have invaded the tissue. Antibiotics agar might be worth a try, if you are able to get it from somewhere.
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roflcopterg
Stranger

Registered: 07/05/22
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Re: Using H2O2 to sterilize tissue before cloning..... [Re: Mushroplex]
#28116682 - 12/29/22 06:46 PM (1 year, 29 days ago) |
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Threads 16 years old lol
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