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Offlinedegs516
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what am i doing wrong??
    #5418327 - 03/19/06 10:20 AM (18 years, 14 days ago)

ok ...ive been doing a g2g transfer in a glove box this is my 2nd time around doing the transfer ...the first time i did it almost all jars got contaminated after the transfer... i back tracked and thought of what i did wrong and did it again and the same thing happened all jars except one are contaminated..
This is what ive been doing

i used Doc's WBS tek for the inoculation and got really good results i got all 4 of my quart jars to fully colonize

I wanted to do a g2g transfer with my jars and get 8 more jars..

The first time i did it i used a glove box.. sprayed the inside with lysol.. waited only like 2-5 minutes.. then did the transfer with all sterile equipement. They all got contaminated with mean green

The second time around i figured i just didnt wait long enough after spraying and that was the problem so when i did it again I decided to use Isopropyl alcohol (because its highly effective in killing bacteria and evaporates very fast). I wiped down the whole box with the Iso sterilized the spoon with a lighter ,wiped it down with iso and put everything in the box.. while in the box i sprayed a couple of sprays with the iso waited 15-20 min. the did the transfer ...and all but 1 got contaminated with mean green.

I thought i took every precaution when doing this and im still getting really high contam. rates

heres a pic



They all mostly have green stuff on the very top of the casing

Does anyone know what im doing wrong??
and can i scoop the green off and try and save it??

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Invisibleagar
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Re: what am i doing wrong?? [Re: degs516]
    #5418424 - 03/19/06 11:02 AM (18 years, 14 days ago)

Did you wipe down the jars - with a sanitizing agent?

Was the mother jar 110% colonized.

Was EVERYTHING in the GB sanitized.

http://www.shroomery.org/forums/showflat.php/Number/1348626#Post1348626

G2G means Grain to Grain transfers.
Once you have learned to prepare sterilized spawn pint or quart jars of bird seed, rye or grains & fully colonize them with mycelium. You can easily propagate a single colonized quart jar of that into about 20 more via G2G transfers.
Spawn rates ranging from 1:10 to 1:40 are being used, with 1:40 being "on the verge of being unsuccessful" [Stamets, GGMM 2000].
Jars propagated via G2G transfers generally colonize under optimal conditions 100% in 10 to 14 days . Given that, spores do not have to germinate, as what is transferred to freshly prepared jars is active mycelium on fully colonized seed or grains.

The method is to prepare fresh jars, just as you would to inoculate via a spore syringe (soak seed, rinse, drain, load, apply filter disk & PC). Excepting, rather than inoculate the fresh jars with a syringe. You transfer grain from a colonized jar to fresh uncolonized jars. The procedure is simple & only requires common sense, minimal preparation, a long stout clean stainless steel spoon & the cleanest personal hygiene and the smallest uncarpeted working place you can muster.

Prepare the smallest cleanest uncarpeted room you have (generally, a bathroom). In the following manner. Clean the room as best you can, getting rid of any dirt, dust, mold or mildew. Remove any cloth hanging anywhere. Spray Lysol on everything, everywhere & wipe it down. If you have any HEPA type air filter unit? Place it in the room & run it for at least 1 hour. Running a HEPA is preferable, but, if you don't have one. You can usually manage without it

Wipe your fully colonized jar of grain & your fresh jars down, with a clean Lysol sprayed rag. Place those in the room, on the counter top, or whatever flat working surface you intend to use. Wear freshly laundered clean cloths. If you have a face mask (preferable) wear it. If you have a shower cap to cover your hair (preferable), wear it. Enter the room, spray Lysol around (again) run the HEPA for a few minutes (if you have one). Then, turn it off. Spray your hands & arms with Lysol & wipe dry.

Unscrew the lid off the colonized jar. Leave the internal filter disk or filter material in place covering the content. Unscrew the lid on a fresh jar, leaving the internal filter material in place. Remove the filter material from the colonized jar & dig up about ? of content, as it will be colonized into a solid mass. Spoon out 2 table spoons full & transfer them to the uncolonized jar, by lifting it's filter up & spooning them in. Replace the filter material on the fresh jar IMMEDIATELY after spooning in the colonized material.

Repeat this same process as many times as you have fresh jars to transfer to. Once done. Screw the fresh jars lids on tight. Cover the outside of the lids of the fresh jars with a double layer of alcohol swabbed coffee filters & rubber band them down. Shake each fresh jar to spread the colonized material throughout it. Place your fresh jars in a dry, dark, warm place (preferably between 78 & 84 F), and allow them to colonize in peace & quiet. G2G transfer & shaking jars batters the mycelium. It takes it a day or 3 recuperate from that shock. There is no need to shake G2G jars more than once. As, doing so will only slow colonization, rather than speed it up.


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Offlinedegs516
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Re: what am i doing wrong?? [Re: agar]
    #5418471 - 03/19/06 11:17 AM (18 years, 14 days ago)

Yeh i wiped down the jars
and everyhting was fully sanitized

The mother jar probably was about 100% colonized id say 95%..but it wasnt getting any more colonized

The mother jar was left in the fridge in a black plastic bag for about a month before the transfer..maybe that had something to do with it

Ive read that somewhere b4 ..but i thought it would be easier to use a glovebox instead of a whole room

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Invisibleagar
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Re: what am i doing wrong?? [Re: degs516]
    #5418506 - 03/19/06 11:31 AM (18 years, 14 days ago)

I use a GB now.
But, before I built one.
I did 100+ G2G's in a sanitized bathroom.
Contam rate was about 2%.


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Offlinedegs516
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Re: what am i doing wrong?? [Re: agar]
    #5418595 - 03/19/06 12:03 PM (18 years, 14 days ago)

if u dont mind me asking ...what is your procedure for using a glove box ..and is it better than the sterile room??

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Offlinedegs516
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Re: what am i doing wrong?? [Re: degs516]
    #5418624 - 03/19/06 12:10 PM (18 years, 14 days ago)

and can i save my jars from mean green ...or should i chuck 'em and start over

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