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OfflineChowder
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Registered: 10/26/09
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What have I done wrong? (100% contamination)
    #15190517 - 10/07/11 07:56 AM (13 years, 3 months ago)

Hello guys,

Coming back to this forum like I'm calling 911 for help.

This is my second grow after a successful first try: 11 jars, 0% contamination, and 45 cracker dry grams. (PF, fruiting chamber).

Now, I have tried a more courageous attempt, with more jars, and got 100% contamination. These are my conditions:
-PF verm+BRF mix
-cooked 90 minutes
-the spore prints: alcohol wiped foil, sealed in several ziplocs.
-clean syringes, cooked for 30-40 minutes.
-innoc in home-made glove box with 2 hand holes
-red hot needle after each jar + alcohol wipe.

Did I leave anything out?
Also, what should I do with the contaminated jars? Can I just empty them out, clean them thoroughly and cook them again for longer?

Thanks guys! Peace out.


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OfflineEvil Toadstool
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Registered: 10/04/11
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Re: What have I done wrong? (100% contamination) [Re: Chowder]
    #15190574 - 10/07/11 08:26 AM (13 years, 3 months ago)

Did your jars contaminate or not germinate at all?

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OfflineChowder
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Re: What have I done wrong? (100% contamination) [Re: Evil Toadstool]
    #15190578 - 10/07/11 08:27 AM (13 years, 3 months ago)

Green coloured mould contamination.


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Invisibleawakendone
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Re: What have I done wrong? (100% contamination) [Re: Chowder]
    #15190598 - 10/07/11 08:33 AM (13 years, 3 months ago)

always sterilize your jars again, before removing the bad cakes from them.

did the contams appear where you injected the spores, or were they at random places on the cakes?

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OfflineChowder
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Re: What have I done wrong? (100% contamination) [Re: awakendone]
    #15190609 - 10/07/11 08:37 AM (13 years, 3 months ago)

Yes, contams have appeared where the syringe injected.. :-s infected needles?


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OfflineEvil Toadstool
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Re: What have I done wrong? (100% contamination) [Re: Chowder]
    #15190620 - 10/07/11 08:40 AM (13 years, 3 months ago)

Most likely a bad print was used. It can be hard to keep a cap 100% clean before and while it drops spores. And yes, you can empty jars and re-sterilize them to be reused.

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InvisibleHesy-Ra
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Posts: 254
Loc: Mars
Re: What have I done wrong? (100% contamination) [Re: awakendone]
    #15190656 - 10/07/11 08:50 AM (13 years, 3 months ago)

Quote:

awakendone said:
always sterilize your jars again, before removing the bad cakes from them




Do this!!! This is for your health and to prevent your whole living area from being contaminated. After make sure to empty them outside. I would rinse them outside also then throw them right in the dishwasher.


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"We must capture a live earthling, K-9, and take him back with us to Mars. Isn't that a nice assignment? Hm?"

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OfflineLord_McLovin
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Re: What have I done wrong? (100% contamination) [Re: Chowder]
    #15190761 - 10/07/11 09:17 AM (13 years, 3 months ago)

Quote:

-red hot needle after each jar + alcohol wipe.




Do not wipe you needle with anything after you flame sterilized it. However, I doubt that this is the (main) reason.

Quote:

-the spore prints: alcohol wiped foil, sealed in several ziplocs.




More information please. Such high contamination rates usually occur when the spore print itself was dirty. How did you prepare your syringes?

Quote:

-innoc in home-made glove box with 2 hand holes




Is it a glove box or a still air box? More info.

Quote:

Also, what should I do with the contaminated jars? Can I just empty them out, clean them thoroughly and cook them again for longer?




Try to kill whatever is inside the jar, before opening it.  I PC anything that appears contaminated, especially if I see any kind of contaminant spores.

EDIT: If the contaminant occurs at the inoculation points, your print was dirty. You probably took the print and put the spores into some water, right? This way you made sure the mold spores were in every one of your syringes.
That's why I always put spores onto agar... I don't know why agar seems to be so unpopular here.


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Edited by Lord_McLovin (10/07/11 09:22 AM)

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Invisibleawakendone
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Re: What have I done wrong? (100% contamination) [Re: Chowder]
    #15190840 - 10/07/11 09:41 AM (13 years, 3 months ago)

Quote:

Chowder said:
Yes, contams have appeared where the syringe injected.. :-s infected needles?




i'd say it's definitely a bad syringe. i wouldn't use that one again. make a new one!

weather i made it or bought it, i always test every new syringe before i do anything serious with it. this has saved me from wasting my time and supplies on quite a few occasions. :thumbup:

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OfflineChowder
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Re: What have I done wrong? (100% contamination) [Re: awakendone]
    #15190861 - 10/07/11 09:49 AM (13 years, 3 months ago)

Should I sterilize the prints? Would that kill the spores?


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OfflineLord_McLovin
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Re: What have I done wrong? (100% contamination) [Re: Chowder]
    #15190970 - 10/07/11 10:20 AM (13 years, 3 months ago)

Yeah, that would kill the mold spores as well as the cube spores. :smirk:

You can try to germinate the spores and clean the culture on agar or just use/buy another print.


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Invisibleprismism
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Registered: 05/11/09
Posts: 5,570
Re: What have I done wrong? (100% contamination) [Re: Chowder]
    #15190982 - 10/07/11 10:21 AM (13 years, 3 months ago)

Your solution is quite simple, buy new spores. Preferably a syringe from a sponsor to insure that it's sterile.


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ephemeral anomalous

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OfflineLord_McLovin
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Re: What have I done wrong? (100% contamination) [Re: prismism]
    #15191164 - 10/07/11 11:02 AM (13 years, 3 months ago)

Of course, then you're missing all the agar fun... :smile:


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