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OfflineB.I.O.
old hand
Male User Gallery
Registered: 05/26/99
Posts: 458
Last seen: 1 year, 1 month
Re: Sterile Syringe Problem
    #92319 - 02/07/00 02:54 AM (24 years, 1 month ago)

hard to say via computer....maybe the prints are already contaminated from your growing area...

send me a part of a print and i can tellya....

BiO.


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Anonymous

Re: Sterile Syringe Problem [Re: B.I.O.]
    #92321 - 02/08/00 08:01 AM (24 years, 1 month ago)

Questions:

#1) How are you sterilizing those syringes?
#2) How careful are you?

Hrm... never did the oven tek, I don't think it's all that great of a tek for making syringes, but is "okay" for innocing jars.

Bleach down room works well for me. As BIO stated it's possible that your print is f*cked. Everything that touches the print must be sterile. That means EVERYTHING you use goes in the pressure cooker before you use it. Bleach down your room, put in the pcooker right after your done cooking, re-bleach the room after about an hour, and then get to work.

This is pretty insane, and I don't do this all the time, just when I screw up a couple times in a row. hehe, but this only happens when I get really sloppy, or got a bad print. (This has happened to me twice..., they were outdoor ones so it wasn't really a bad print per se)


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OfflineCrobih
rap-cord
Registered: 11/03/98
Posts: 2,015
Loc: cave
Last seen: 11 years, 6 months
Re: Sterile Syringe Problem [Re: B.I.O.]
    #92322 - 02/08/00 08:47 AM (24 years, 1 month ago)

Maybe, you should make really dens spore syringe (put much as you can spores), leave it few days to germinate(?), shake them sometimes ,then inoculate in cakes (dont be cheap), keep cakes in cool for 3-4 days and then put them in 80F. It worked for me ,
and I had very filthy sporeprins fro mthe nature (not just once).

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Anonymous

Re: Sterile Syringe Problem [Re: B.I.O.]
    #92323 - 02/09/00 12:47 PM (24 years, 1 month ago)

Would adding peroxide to the water help in preventing contamination? I know that spores and mycelium will tolerate peroxide, and that most bacteria won't. Anyone try this?

------------------
I feel the urge to take my fist, and violate your every orifice!

Peace & Love, ya'll


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OfflineCrobih
rap-cord
Registered: 11/03/98
Posts: 2,015
Loc: cave
Last seen: 11 years, 6 months
Re: Sterile Syringe Problem [Re: B.I.O.]
    #92324 - 02/09/00 02:11 PM (24 years, 1 month ago)

Spores won+t tolrate it

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OfflinePipes of Pan
journeyman
Registered: 08/16/99
Posts: 18
Last seen: 4 months, 7 days
Re: Sterile Syringe Problem [Re: B.I.O.]
    #92325 - 02/09/00 05:04 PM (24 years, 1 month ago)

The problem with contaminated spore prints makes a good case for starting on agar. Even if your print is contaminated, you can eventually isolate the clean mycelium if you start it on agar medium. Sure, more work, but once you have your culture started and clean, you have the culture.

Pipes

------------------
Pipes of Pan


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Anonymous

Re: Sterile Syringe Problem [Re: B.I.O.]
    #92326 - 02/10/00 06:54 AM (24 years, 1 month ago)

Okay ladies and gents... time for an ootw special. Isolating on agar from nasty critters.

First of all. You need to make sure your print is nice and dry. Stick it in a dessicant chamber for a couple days. This should any "living" things, but not spores, etc.

Then put it on some agar. Let it grow out a bit, (once you see both mycelia and contams) and if you see some contams, take a syringe of 100% (yes thats 100) h202. We're only talking 2cc's. And zap the bastards with it. Note that this procedure might have to be repeated several times.

Does it work? Not always. But I isolated the strain of Agrigarius Bisporus that I use for my yummy omellettes this way. Turned out to be one of the most vigorous strains in my collection of edibles. (Correction, the most vigorous)

I did however forever lose a strain of Ps. Cyans from Canada because this tek didn't work. I got some other suggestions, but alas it was too late, all the syringe had been used.

By the way, another great tek is growing some out on agar, and then flopping another agar gel on top of the contammed section. You see mycelia will grow up through agar, but almost nothing else will! :smile: Also use of wood chunks on top of the agar, there is an involved tek that stamets discussed, but I can't remember the details. If your interested email me.


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InvisiblePrellgott
addict
Registered: 02/08/00
Posts: 383
Re: Sterile Syringe Problem [Re: B.I.O.]
    #92327 - 02/09/00 10:10 PM (24 years, 1 month ago)

...good idea but the classic way is just transfer some clean mycelium of the agar plate to a new one and agar and again untill it is clean..
anyway you should try mycelium transfer and Old Lazlo`s Mycelium syringe Tek....it`s better for beginners
(check that thread in basic cultivation)

[This message has been edited by Prellgott (edited February 10, 2000).]



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