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Offlineinsutama
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Starting over
    #25467548 - 09/17/18 05:52 PM (2 years, 7 months ago)

So at this point im pretty sure my tubs are contaminated there is a white cauliflower looking growth on them. This time im gonna try starting with agar. I have 2 no pour mini rounds in my SAB along with a spore print and a inoculating loop im letting them sit in there for a good hour before i attempt to inoculate my agar. I just wanted to kinda run threw my procedure and hopfully somone can tell me if im doing it wrong or right i am quite new to working in a SAB and with agar.

First wash my hands and arms with anti bacterial soap then put on gloves and proceed to unwrap the foil that my inoculating loop is in inside the SAB.

flame sterilize my loop and cool it by wiping it on a alcohol soaked paper towel

open my spore print in the SAB then with my loop scrape some of the spores.

crack the lid on my mini round and run my loop in a zig zag motion over the agar and quickly close the lid on the mini round.

then flame the loops again and repeat with the other rounds.

Am i missing anything ?
is it okay to leave the print open in the SAB or should i be just pealing back the foil on the print a tiny bit then folding it closed again in between agar containers ?

any advice on how to work in my SAB to prevent contams would be greatly appreciated. I just spent over a month with my last attempt and had nothing but failure. Its been almost 5 years since my last grow and i always just did spore syringe into rye jars and never had a issue however many times on my second flush i would get mold form in my tubs which leads me to believe i never had a clean culture to begin with in my jars hence why i want to get into agar work.


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OnlineGrimsweeper
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Re: Starting over [Re: insutama]
    #25467555 - 09/17/18 05:55 PM (2 years, 7 months ago)

First thing you're doing wrong is don't cool your Loop on an alcohol soaked paper towel. Cool it on the agar. You're wiping out any sterilization you just did with the flame


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Offlineinsutama
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Re: Starting over [Re: Grimsweeper]
    #25467614 - 09/17/18 06:18 PM (2 years, 7 months ago)

okay makes sense
Can i leave the lid of the agar while i cool the loop and then inoculate it ? or should i just crack the lid in between


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InvisiblebodhisattaM
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Re: Starting over [Re: insutama]
    #25467617 - 09/17/18 06:18 PM (2 years, 7 months ago)

You cant put the lid down on anything but its own dish that it came from


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Offlineinsutama
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Re: Starting over [Re: bodhisatta]
    #25467646 - 09/17/18 06:29 PM (2 years, 7 months ago)

okay


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OfflineFuturepastnpresent
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Re: Starting over [Re: insutama]
    #25467700 - 09/17/18 06:50 PM (2 years, 7 months ago)

Go very slow.


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Offlineinsutama
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Re: Starting over [Re: Futurepastnpresent]
    #25467747 - 09/17/18 07:05 PM (2 years, 7 months ago)

okay thanks for the tips im gonna go attemp this


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