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 Zen Peddler
 
Registered: 06/18/01
Posts: 6,379
Loc: orbit
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 Cloning without agar?? 
#535116 - 01/29/02 04:10 PM (22 years, 2 months ago) |
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Couldnt I make a sterile solution of say 1ml of malt extract/honey and 2-3mls of h202 to 100mls of water and into this, add a sterile (soaked in h202) piece of a fruit i wish to clone - then if mycelia grows from this i could extract it into a syringe??
Due to the fact that there arent any spores to germinate i could even up the h202 to make sure it is sterile...
Any thoughts??
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CLuB99
lost somewhere in time and space
Registered: 10/26/99
Posts: 1,316
Loc: my mind
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 Re: Cloning without agar?? [Re: Zen Peddler]
#535122 - 01/29/02 04:20 PM (22 years, 2 months ago) |
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it works....go for it!
mmm why don't you prepare some peroxiagar and clone the mushroom with it???
works way better than honeywater
just my 2 eurocents...
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Zen Peddler
Registered: 06/18/01
Posts: 6,379
Loc: orbit
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 Re: Cloning without agar?? [Re: CLuB99]
#535125 - 01/29/02 04:24 PM (22 years, 2 months ago) |
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I dont have the supplies at the moment - more just curious - anyway, thanx for the reply
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Champion des Champignons
long standing member;)
Registered: 07/26/00
Posts: 2,680
Loc: Alba
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Re: Cloning without agar?? [Re: Zen Peddler]
#535157 - 01/29/02 05:04 PM (22 years, 2 months ago) |
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I've had partial success with just chopping up a mushroom stem and mixing it up with sterilised grain and 10ml of h202. It contaminated just before 100% colonisation, but I was very sloppy about it, no glovebox, didn't even sterilise the knife I chopped the shroom up with. I would imagine if you weren't as slapdash as I was you'd have a good chance of success.
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hmmm........
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 Lizard King
King Lizard
Registered: 10/03/99
Posts: 1,998
Loc: GA
Last seen: 17 years, 7 months
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I use a BRF paste all the time for cloning, just add water to brf until you get an almost pourable consistency and wallah. I usually make it a little on the runny side because it tends to loose some of its moisture during sterilization, if not enough water is used to make the paste it will harden a bit and make colonization slow. Other than that it works great for clones or regular spore work, even squirted syringes on the stuff before and had great sucsess. From my observations I concluded that brf paste is harder to contaminate than agar and it is better than agar for clones because of the dirty nature of cloning, especially wild fruits.
Try it, it works 
LK,
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Zen Peddler
Registered: 06/18/01
Posts: 6,379
Loc: orbit
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 Re: Cloning without agar?? [Re: Lizard King]
#535616 - 01/30/02 03:18 AM (22 years, 2 months ago) |
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thanx lizardking - would you suggest adding h202?? And how much would suggest - especially if your using wild fruit-bodies? And do you know if clones produce spores??
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Lizard King
King Lizard
Registered: 10/03/99
Posts: 1,998
Loc: GA
Last seen: 17 years, 7 months
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Re: Cloning without agar?? [Re: Zen Peddler]
#536198 - 01/30/02 05:45 PM (22 years, 2 months ago) |
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Yes, adding some h202 will help to clean the clone tissue. I'm not sure if you already know this, but you can't add h2o2 to anything that will go in the pressure cooker as it will be broken down into regular h20(water). So you will need to add it to the sureface of the brf paste after you sterilize it. The way I did it and others have told me to do it, it just run your tissue through a coffee grinder into a cup of water with peroxide mixed with it, then suck this up into a syringe and inoculate you brf paste. I guess you could also inject a small amount of peroxide on the surface of the brf paste then clean your tissue by dipping it in peroxide and then drop it on the brf paste. I dunno, I'm sure there are many ways to do it I would definitely use peroxide when cloning though, especially wild clones.
BTW: In case your wondering because I didn't post it, you should pour the brf paste the same depth as you pour agar agar, it works in petris or jars just the same.
LK,
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SThomas
enthusiast
Registered: 10/15/01
Posts: 251
Loc: Ottawa, Ontario
Last seen: 21 years, 11 months
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Re: Cloning without agar?? [Re: Lizard King]
#536320 - 01/30/02 08:11 PM (22 years, 2 months ago) |
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Before I bought some agar, I used brf with very good success. I t works great. Now with the agar I mix some grain juice and brf with the agar...after I have sterilized the agar..I add the H202...zero contams so far. I love agar way less problems than just inoculating jars with sporeprints. And a much faster colonization time when inoculating a substrate with it. I just inoculated 6 jars of birdseed with agar and 1.5 days later I have 10% colonization...should happen real fast now. But I guess the whole point is that brf works great too...and I swear by H2o2...has really made things work better. When I inocluate the jars I just use the open oven tek...works excellent, without having to build a flow hood or glove box...Cheers good luck
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And that is all the news that's fit to print
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