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Cloning


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#1 MurCurY

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Posted 28 February 2007 - 12:10 AM

pics from my clonging process.

Pleurotus eryngii

http://mycotopia.net...=1&d=1172640578




I cut the bottom off....then use the flamed knife to make a 1/16" deep cut across the stem on both sides.

http://mycotopia.net...=1&d=1172640578
http://mycotopia.net...=1&d=1172640578


Then break it open to properly pull some clean tissue.
http://mycotopia.net...=1&d=1172640578

Using my specimen needle I fish some tissue from the inside of the stem taking care not to touch it. I also flame the needle and dip it in peroxide before fishing out new pieces. for the P. eryngii I made 4 plates so...4 pieces.
http://mycotopia.net...=1&d=1172640578


a sample of tissue.
http://mycotopia.net...=1&d=1172640578


Only need a small piece.
http://mycotopia.net...=1&d=1172640578

I then dip the samples in peroxide for 30 seconds or so. Then plate them in the center of agar poured into half pint ball jars. I poured these thick...I'll make them thinner next time.


Pic of plated sample.
http://mycotopia.net...=1&d=1172640578

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#2 message2harry

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Posted 28 February 2007 - 12:47 AM

wow awesome. I plan on cloning in LCs pretty soon. awesome job and great pics!

#3 golly

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Posted 28 February 2007 - 08:01 AM

Great pix Mon...!...Good luck with those...

#4 siam_jim

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Posted 28 February 2007 - 09:32 AM

murc nice intro dude, great pictorial too, what type of agar (pda?) are ya using there?

siam

#5 fahtster

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Posted 28 February 2007 - 09:40 AM

nice! :)

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#6 dial8

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Posted 28 February 2007 - 10:43 AM

Very nice. Looks real good.
So are you working in open air or are those pics simply for illustration?

#7 Bobcat

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Posted 28 February 2007 - 01:01 PM

:bow:

#8 MurCurY

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Posted 28 February 2007 - 01:13 PM

I'm using MEA. Brew shop bought malt and korean store "telephone brand agar"

I poured my jars then pc'ed them. I do work in open air. I'm building a flow hood with a uvc light and a hepa after i move in the next couple weeks. so that will help contam rate if any. I've had good luck with open air before. guess we'll see.

#9 Bobcat

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Posted 28 February 2007 - 01:21 PM

What do you think your contam rate is in the open, doing a project like this? Percentage wise? How are you planning to implement the uv light? On incoming air?

I've used jars, but after switching to plates there is no going back for me, personally. Have you worked with them yet? But your system looks real good as is!

#10 MurCurY

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Posted 28 February 2007 - 01:55 PM

i've used plates. i have a few sleaves left. decided to try jars cause you can pour then sterilize them. that's where i'm getting most of my contams i think. on actually pouring my dishes. when i move and have my box setup i'll try pouring some plates in the flowhood and see contam rate. I'll let you know what my contam rate for this project is....this is the first time trying it this way. Even if it doesn't work out as planned.....at least i learned something ;)




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