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cloning oyster tissue to grain


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#1 mydarling

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Posted 30 October 2010 - 12:42 PM

when cloning mushrooms via tissue sample into jar of grain ....

is it best to bury the tissue deep within the grain?
or can it be left on top of the grain?

a few nights ago i took some tissue samples and simply placed them on top of my popcorn, so that i could see what's going on as it starts to produce myc ... however nothing has happened yet, i.e. tissue is still sitting atop the popcorn looking exactly as it did when i put it there.

would it be better to shake up the jars so the tissue is buried within the grain, not sitting exposed on the top? would this promote faster myc growth?

thanks!

p.s. the tissue was sampled from oysters, if that makes any difference, but i figured this question was general enough to go in the main fungi forum.

Edited by mydarling, 31 October 2010 - 10:39 PM.
title

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#2 mydarling

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Posted 30 October 2010 - 12:50 PM

ok, so i just checked them again, and since i checked last night, a light fuzzy myc layer has formed on the tissue chunks, so i guess it is working after all :teeth: but the question still stands, would it be BETTER if i shook them up and buried the tissue?
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#3 Freaky

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Posted 30 October 2010 - 01:31 PM

I tissue transfer to lc and have never done tissue to grain.

I'd think on top may cause the tissue sample to dry out a little holding off a good start to colonization. So maybe shaking to mix the tissue would help a little.

When I've used agar wedges to grain I've shaken the jars to distribute the grain.

#4 mydarling

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Posted 30 October 2010 - 01:44 PM

ok, thanks freaky! :) will shake 'em up!

i did 3 tissue transfers to LC also. i'm not really sure if my LC's are going to turn out well, though, so i did a bunch of grain to be safe. we shall see!!! i'm very excited to see beginnings of fuzzy myc. :thumbup:

#5 Freaky

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Posted 30 October 2010 - 01:52 PM

Awesome MD! It's good you spread out your distribution of tissue samples to a variety of sources. That way you can see which will be preferable for you.

What flavor oysters are you working with?

#6 stellarj

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Posted 30 October 2010 - 03:12 PM

they say that if you get the myc to start growing from the middle the outside layer will all be the same age and growing fast when its getting done rather than having old growth on top newer in the middle and new growth trying to get thru the bottom when it is fully colonized. i am watching shitake colonize some jars of wooden dowels right now. good luck. j

#7 mydarling

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Posted 30 October 2010 - 04:07 PM

freaky, i'm not sure what kind of oysters they are. i just bought them from the grocery store and all they said was "oysters"! look kinda like pearl to me, though.

IMG_0043-resize600.JPG IMG_0044-resize600.JPG


thanks for that info, stellarj! i have shaken them all up now, i'm not sure if the tissue ended up in the "middle" per se, but it's buried inside somewhere and i can't see it.

but i did snap a few pics of the DA FUZZ before doing so :amazed:

IMG_0095-resize600.JPG IMG_0098-resize600.JPG IMG_0099-resize600.JPG

oh mycology, how i love you :heartbeat :heartbeat :heartbeat


Edited by Sidestreet, 13 January 2017 - 06:22 AM.


#8 roc

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Posted 30 October 2010 - 04:19 PM

Looking good...

#9 mydarling

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Posted 30 October 2010 - 04:27 PM

thanks for the vote of confidence roc!

heh i guess i better move this to the oyster forum now, since i've got oyster pics and whatnot.

#10 Freaky

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Posted 30 October 2010 - 05:43 PM

Looks like Pearls to me as well. Nice start MD, and stellarj made an interesting point I didn't think of by leaving it on top with the mycelium and the age as growth spreads. Thats good to know.

I can't wait to watch this progress for you.

I :heartbeat mycology too
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#11 Nobody

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Posted 30 October 2010 - 05:55 PM

We have lift off! :eusa_clap

#12 copelandiaKidd

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Posted 31 October 2010 - 08:05 AM

thats exactly what i did with my oysters..... store bought, tissue to grain..... worked just fine!!!!!

#13 kpop

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Posted 31 October 2010 - 08:49 AM

so could this method be done the same way for cubies and exotics

#14 Freaky

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Posted 31 October 2010 - 09:25 AM

so could this method be done the same way for cubies and exotics



Yes.

#15 kpop

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Posted 31 October 2010 - 05:22 PM

nice, keep us up to date on how its going copelandiaKidd
looking good.
:)


#16 mydarling

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Posted 31 October 2010 - 10:25 PM

nice, keep us up to date on how its going copelandiaKidd
looking good.
:)


hey now, cope has his own thread (http://mycotopia.net...clone-grow.html), this one's mine :lol:

oh yeah, and cope i've been following your oyster clone grow and loving it! that was part of what inspired me to get a project going. ;) i didn't feel like making a blender slurry jar though, so i just did it real quick and simple. chopped off the cap and base of stem, soaked the stem portion in 3% hydrogen peroxide (H2O2) for a couple mins, then used a sterilized scalpel to slice down the stem and peel back the outer layer. the actual clone tissue was taken from a deeper layer inside the stem. i dipped the inner tissue chunks in H202 for about 20-30 seconds and then dropped them into PCed popcorn jars. all done inside a glovebox, of course :)

maybe i should turn this into a real grow log ?
i have taken a shitload of pics.
if there is interest let me know :rasta:

#17 shroom_seeker

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Posted 31 October 2010 - 10:27 PM

Yes, we are interested, make it official MD! :headbang:

Myc looks good, can't wait to see some oysters. :loveeyes:

#18 mydarling

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Posted 31 October 2010 - 10:38 PM

Yes, we are interested, make it official MD! :headbang:


haha, ok :hugs: let me go resize some pics and whatnot. ;)
i guess i better change the thread title too!
good thing i'm a mod and can edit my own threads :eusa_danc hehe

#19 mydarling

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Posted 01 November 2010 - 12:27 AM

ok let's see here, i just went through all the photos and tried to remember what is what ;)

so i made some fancy lids with 4 high temp RTV silicone self-healing injection ports , and a double-washer micropore tape FAE hole in the center :rasta: -- sort of modified BB lid tek, he uses tyvek but i used micropore instead. and the holes are different size i think.
1. punch 5 holes w/ nail and hammer, wiggle nail in hole to widen holes slightly
2. drill outer 4 holes to 1/4" diameter (for injection ports - 4 holes means these lids can be used for cakes or grains as desired)
3. drill center hole to 5/16" diameter (for FAE)
4. apply blob of RTV silicone to 4 holes on outside of lid. swirl around to get out any air bubbles. flip lid over and apply another blob on the inner surface of the holes to make complete ports.
5. apply RTV silicone around perimeter of the center hole, add a 1/4" zinc washer, and press slightly to squish out air bubbles. flip lid over and apply another washer on the inner lid surface.
6. allow to set for 24 hours. apply micropore tape to both inner & outer washers. this creates a "dead space" between the two tape layers and is another barrier against contam, it also prevents any fluid from the grains wicking out.
oh, also did some basic polyfil lids (5/16" center hole stuffed w/ poly).
lids-16-resize1000.JPG

did up a bunch of PCORN (pints & quarts) by turkeyranch's tek, see http://mycotopia.net...-pictorial.html
post-pc-14hr-02-resize1000.JPG

i let them sit for 4 days (some at incubation temp, some in fridge) to make sure grain texture & sterilization was fine. they still looked good so i decided to inoculate :thumbup:
post-pc-4d-01-resize1000.JPG

i also did up 3 LC's:
1. 1 tsp clover honey + 1 tsp light karo + 800mL distilled water (approx measurements)
3. loaded 250mL LC mix into each of 3 pint jars, using lids w/ 2 self healing silicone ports: one for injecting/extracting & one for airport syringe
2. LC's and airport syringes PC'ed at 15 PSI for 30 mins
3. allowed to cool in PC overnight til vacuum seal formed, then airports inserted into 1 port
lc-oys-day00-01-resize1000.JPG

the store bought oysters were cloned by the peroxide method i mentioned above. here are some pics from that. (i did 1 jar outside GB in open air so i could take pics, and also just to see if i could get a clean jar from it :D)

cap & base chopped off w/ sterilized scalpel
osyters-10-resize1000.JPG

stems put in bowl of H202
osyters-09-resize1000.JPG

outer layer peeled off w/ scalpel (sorry could not get a good pic)

inner tissue removed with scalpel
oysters-12-resize1000.JPG

placed again into H2O2 for a few seconds
oysters-13-resize1000.JPG

some tissue chunks went into grain jars, some went into LCs
lc-oys-day00-03-resize1000.JPG

the fuzzy myc pics in the post above were 2 days after inoculation.

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#20 Nobody

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Posted 01 November 2010 - 07:06 PM

Nice roundup of your work, looks like you're well on your way. Here's to some fatty flushes of oysters! :headbang:

How are you planning to use the grain spawn?




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