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dry tissue cloning ?


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#1 sandman

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Posted 16 March 2005 - 12:02 PM

I about to do some agar work on a peice of dried shroom as i got nothing else lol. I hear to mix the agar a little "wetter" (less agar to water is what i did). What else should I do? The info i found in the archives was a little slim.

Is it a good idea to dump the peice in h202? I dont get along with h202 very well but I mean this peice of mushroom has been sitting in a jar of dissicant for 6 months with a bunch of coir and vermiculite from the harvest in there with it.

Is a peice the size of a grain of rize about right?

BTW im not using any kind of crazy h202 agar or antibiotic agar. I know that contams will grow too, but was hoping to either cut away the good stuff or do the "hot agar on top" possibly with antibiotic.


I know roger knows about this!

#2 Guest_pissybee_*

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Posted 16 March 2005 - 02:27 PM

Might soak the piece in water first, before trying and I'd use that antibacterial agar. Prolly have to do some cleaning up, maybe some peroxidated agar? Never tried this so I am just throwing out some suggestions, rodger is the one to really ask about this, though...

#3 sandman

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Posted 16 March 2005 - 02:53 PM

also, is it cool to re-PC some mixed up agar? I have a quart jar half full that I put in the fridge after pouring some plates (PCd for like 30 minutes) a few days ago. Can I PC it again and pour again, or will this be too much cooking for it? Do I need to mix up some new stuff?

#4 1dumbmyco

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Posted 16 March 2005 - 06:25 PM

Hell my friend got away with just a karo jar, no agar, no PC...just hips microwave tek and chunk of dried EQ tissue. It took a while, didn't produce much viable myc, but preserved a strain that had escaped him through inadvertent trades of prints.I had thot hip had marked the thread @rchive but am unsure. Good luck and it does work since fungi are such simple organisms and that is the preferred method of spreading strains in the commercial industry, dry material.

#5 destroy_erase_improve

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Posted 16 March 2005 - 07:43 PM

i dont know much about agar either, although i love to see pics of it and read about it.
maybe drop it in peroxide for a few seconds, let it fizz, then drop that small piece on the agar in front of the flowhood/inside glovebox.

after you get growth cut away any clean myc from contams, and do the same in a new petri.
thats just a guess of what they are going to say to do.

i do remember reading about how you can pour new agar over the contamed agar , and the myc will poke through, you then scrape away the clean myc and put it on a new dish to grow out.

but it really all comes down to someone with first hand experience, and not just a reader like me.
good luck

#6 Guest_Peter Cottontail_*

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Posted 16 March 2005 - 09:02 PM

I would recommend you try to surgically get a piece of dry tissue from the inside of the stem if you can. If not, just snap off a piece of tissue and drop it into the agar. I never dip in peroxide first. Kinda cut and push it in to put a bit more tissue in contact with your agar. It will take about a week for anything to start happening. Sometimes spores that are stuck to the stem will germinate before the dry tissue starts growing again. You'll know this because there will be sectors of different substrains of mycelia growing. If it's a pure clone from your stem, the mycelium will be uniform across the petri dish.

Contamination will definitely grow too, but you're learning agar, so jump right in and transfer your healthy mycelium to a new petri dish. Never try to move contamination away from a dish. When contamination shows up, I shut the flow hood off and work in a glovebox. I don't want my flow hood blowing contam spores everywhere. Make a couple of transfers of healthy mycelium away from the original dish, then do the hot agar tek as a final clean. After that, transfer a wedge to rye grain, and away you go. I've used this tek on two year old dry tissue with success.

#7 destroy_erase_improve

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Posted 16 March 2005 - 09:30 PM

wow
i like the tidbit of info about turning the flowhood off

so when you see contams and myc growing away from it, you move to a glovebox to work . and transfer the seemingly healthy myc from the contamed petri to a new fresh one. how do you prep the glovebox . just bleachwater it down? use an ion generator with it?

this is cool

#8 elixir

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Posted 16 March 2005 - 11:23 PM

A foaf did that karo blender trick and it worked. His jar didn't get a thick mat of mycelium but when he shook it up and then used it to innoc it grew well. He didn't sterilize the needle though so the jars all got contams.

#9 Guest_Peter Cottontail_*

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Posted 17 March 2005 - 01:35 AM

I just wash the glovebox with soap and water before and after each use.

#10 Guest_novak_*

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Posted 08 April 2005 - 03:35 PM

has anyone tried it? cardboard is much easier to deal with then agar. i've seen that it is possible to clone from dried tissue on agar but what about cardboard? anyone up for an experiment?

#11 Soliver1

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Posted 08 April 2005 - 03:38 PM

I wouldn't hold my breath, but give it a shot and post a pic
if it pans out preternaturally.

:)

#12 Lazlo

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Posted 08 April 2005 - 07:13 PM

http://mycotopia.net...read.php?t=3093

#13 Guest_lost_onabbey_rd_*

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Posted 08 April 2005 - 07:28 PM

i've tryed reviving dryed tissue on carboard in a karo solution (card board was colonized then dryed).. after almost 2 months 1 of the 2 karo cultures showed enough growth to try some jars... i have done 2 brf jars that have shown no growth for almost 2 weeks... i'm keeping an eye on them.. but i don't think they are gonna do anything..
after the tissue is dryed it is my impression that it is hard to revive. from what i've heard even reviving dry tissue on agar normaly results in show growth and little vigor.
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#14 Guest_Peter Cottontail_*

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Posted 09 April 2005 - 04:55 AM

Cardboard is not a replacement for agar. It has very little nutrition. Dry tissue grows again very well on agar. It will also have its original vigor from my experience, provided it wasn't suffering from senescence before it was dried. Build a glovebox and learn agar work. It really is the next step.
RR

#15 Guest_novak_*

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Posted 09 April 2005 - 10:13 AM

agar is the next step but it's a tad intimidating.

#16 Guest_lost_onabbey_rd_*

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Posted 09 April 2005 - 09:26 PM

well i stand corrected :)
RR when re-growing live tissue on agar is it nessacary to hydrate the tissue or do you just stick it on the agar still dry?
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#17 highflyer

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Posted 10 April 2005 - 12:31 AM

Id give the tissue a drop of water on the agar if its dry.

#18 Guest_Peter Cottontail_*

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Posted 10 April 2005 - 07:10 AM

Yea, especially if the agar is dry. I put the dry tissue on agar as soon as the plates cool down from pouring while they're still fairly wet. This hydrates the piece of tissue. I just started some two year old dried culture on agar last night, so I'll try to do a photo tek on the process.
RR

#19 MurCurY

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Posted 27 November 2005 - 12:03 PM

I have tons of agar supplies now.
tons of Malt extract and potato dextrose and agar powder.
I want to make up some Malt extract agar. would someone be willing to share a winning recipe? Also...i was looking through the archives and found an entry for someone growing dried tissue on agar....how would i do this? I have a piece of cap i saved from some shrooms i had about a year ago...will this grow on agar? will i have to hydrate it....

#20 MurCurY

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Posted 27 November 2005 - 12:40 PM

here's one of the posts...
http://www.mycotopia...html?1086014580




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