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OfflineAFOD
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Cloning wild Mushrooms
    #6826454 - 04/24/07 09:01 AM (14 years, 18 days ago)

Found more oysters in the mountains. Look almost identical to the ones in the back yard, except these appear to be more tender. I'm guessing maybe it's a sub-strain or something. Anywhat, I have prepared several substrates for the transfer of live tissue. I know that all my other transfers were contaminated with green mold (which had to have been growing on the stem, because I used H202 in the LC) I am wondering if one could possibly wash the stem of the specimen to be cloned (say with watered down Lysol, then H202, then rinsed well with clean water) without killing the sample.
By watered down lysol I mean I found lysol in a concentrated form and would mix it lighter than reccomended by MFG.
My goal is to cut down the chances of transferring live mold tissue with the desired tissue.


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OfflineJeremy_Davis
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Re: Cloning wild Mushrooms [Re: AFOD]
    #6827355 - 04/24/07 02:14 PM (14 years, 17 days ago)

I sometimes wipe down mushrooms to be cloned with an alcohol pad.

It's really hit-or-miss when you clone directly to LC. If you use agar plates (agar can be bought at almost every oriental grocery store for about $1-2 for 25 grams), then even if it is contaminated, you can do your best to isolate away from the contamination. In an LC, the contam is EVERYWHERE...

just my thoughts...


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OfflineRogerRabbitM
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Re: Cloning wild Mushrooms [Re: Jeremy_Davis]
    #6827678 - 04/24/07 03:44 PM (14 years, 17 days ago)

You need to learn agar work. Wash wild mushroom tissue with iodine, and then place on agar. Iodine will kill most of the bacteria, and then you make a series of transfers to get the mushroom mycelium to outrun the molds that are on every outdoor mushroom sample.
RR


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Invisiblethallus
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Re: Cloning wild Mushrooms [Re: RogerRabbit]
    #6827888 - 04/24/07 04:40 PM (14 years, 17 days ago)

Quote:

RogerRabbit said:
Wash wild mushroom tissue with iodine




How long do you wash for?


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OfflineRogerRabbitM
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Re: Cloning wild Mushrooms [Re: thallus]
    #6828133 - 04/24/07 06:03 PM (14 years, 17 days ago)

Not long. I rinse them with iodine by holding them over a sink so the iodine can run off the mushroom and down the drain. I then dip each piece of tissue in iodine after cutting it out before placing on agar. This helps to prevent bacteria from taking over your plate before the mushroom mycelium has a chance to get started.

10% iodine is what I use because it's what the local pharmacy carries and it works well, so I know it's safe. If yours is stronger than that, you might want to dilute it with distilled water.
RR


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semper in excretia sumus solim profundum variat

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OfflineCryogenicz
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Re: Cloning wild Mushrooms [Re: RogerRabbit]
    #6828238 - 04/24/07 06:29 PM (14 years, 17 days ago)

You can also make a sanitizing solution from idophor used in home brewing.

-Graham


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OfflineAFOD
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Re: Cloning wild Mushrooms [Re: Cryogenicz]
    #6829011 - 04/24/07 09:54 PM (14 years, 17 days ago)

We would really like to use agar, but live in a very rural area and there are not any ethnic stores whatsoever within 150 miles. Bacteria is not our biggest concern because we use H202 in our live cultures and transfers, which has proven to be very effective, except in cases of live mold. Since the mold growing on the wild specimens isn't even visible to the naked eye, we're thinking it could be at least partially eliminated by light use of lysol (which kills fungus) on the exterior of the sample. Then H202 to ensure that no bacteria or mold spores are able to germinate.
I guess we'll just try it. We are going to take prints from the mushrooms and save them for a time when we can get agar.
BTW, what is idophor?
The second mushroom colony we found look very tender and delicious. We would really like to get some growing.
Y'know what sucks? We purchased pre-mixed agar from the sporeworks in thier beginners culturing kit. Then during our recent move the box got lost.
I am starting to doubt that we'll be able to successfully clone without the use of real agar. So, like I say. we'll collect spores and use them once we have some agar. Thanks all.
Hey! What about letting the mushrooms sporulate into jars of the rye flour gravy agar sub we made. If enough spores are dropped, it seems likely that they would colonize quickly and not give contams a chance.


Edited by AFOD (04/24/07 10:03 PM)


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OfflineRogerRabbitM
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Re: Cloning wild Mushrooms [Re: AFOD]
    #6829193 - 04/24/07 10:28 PM (14 years, 17 days ago)

Peroxide is horrible for cloning because it damages the mushroom mycelium you're trying to clone, setting it back and giving the faster recovering and growing molds a chance to take over. Save your peroxide for combatting dactylium(cobweb) mold. It's of little other use.

You can get premixed agar powder from sporeworks or fungi.com
RR


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semper in excretia sumus solim profundum variat

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Thomas Edison


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OfflineAFOD
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Re: Cloning wild Mushrooms [Re: RogerRabbit]
    #7404908 - 09/13/07 05:58 PM (13 years, 7 months ago)

So RR, is the iodine left on the tissue transfered onto the agar? You said you rinsed it off then dunked it again. I suppose this doesn't affect the agar or culture negatively?


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OfflineAFOD
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Re: Cloning wild Mushrooms [Re: AFOD]
    #7404918 - 09/13/07 06:00 PM (13 years, 7 months ago)

and how affective is H2o2 against growing cobweb mold? I have found nothing on these subjects before. Greatly apprecialte your advice, as always RR you are the MAN!
If you could point me to some links I can read them.


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OfflineRogerRabbitM
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Re: Cloning wild Mushrooms [Re: AFOD]
    #7405030 - 09/13/07 06:32 PM (13 years, 7 months ago)

Peroxide will kill cobweb mold. Use it at 3%, which is the concentration it's normally sold at in the USA.

Rinse with water after the iodine. In fact, I tear a piece of tissue with my hands from the wild mushroom, and rinse it over the kitchen sink with iodine. When I get into the sterile room/glovebox I tear the piece in half, and cut a tiny piece no larger than a grain of rice from the center of the chunk of tissue. Be sure to transfer tissue that has never been exposed to air until you've just tore it open in sterile conditions.

For extremely contaminated outdoor fruits, you can do a series of sterile water baths. Sterilize several jars of distilled water. Drop your tissue into the first jar and shake well. Transfer to the second jar and repeat. Three or four rinses prior to placing on agar will go a long way towards ensuring success. You can put a teaspoon of laundry bleach in each pint jar of water you rinse with. The bleach will help kill off bacteria and mold spores, but leave the mushroom mycelium undamaged.
RR


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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
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Mushrooms, Mycology and Psychedelics >> Gourmet and Medicinal Mushrooms

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