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d0urd3n
Just call me "D"


Registered: 09/15/10
Posts: 5,237
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First attempt at agar felt like a shit show
#20198528 - 06/28/14 11:35 AM (10 years, 6 months ago) |
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I always think I have it all planned out and it will be easy until I actually get my hands in that SAB.
Is there a very detailed explanation of sterile technique for this?
I put my half pints in there, realized I should have wiped them down, took them back out, wiped down, put them back in. I wasn't sure if I should wipe down the foil of the prints, but I figured the inside was just as dirty. Not very easy to open those in an SAB with gloves on. My ghetto inoculation loop I wasn't sure where to put it when it was sterilized while opening the lids/loops. I guess I should have cracked the jar and mostly opened the print and THEN flamed it and went to work. The prints that I used were tiny and light, I tried to kind of scrape them but it was hard with my flimsy loop. I figured it shouldn't matter if I don't see them since they are microscopic, but still.
Should have misted in between each print>agar. Initially I thought it would be fine since I soaped and watered down the inside and outside. I think I will buy an alcohol burner to just reduce one step/chance for contam.
I rubbed it all over the agar to make sure I got the spores off which I realize was probably a bad idea. Definitely need to attach some type of handle to my loop made out of a twisty tie.
I guess all I can do now is wait. Sterile technique really is quite a skill though especially in an SAB I now realize.

At least my agar turned out pretty well. Next time will be smoother. 
edit: if they turn out clean should I still make a transfer just to be sure or should I throw some wedges in grain?
Edited by d0urd3n (06/28/14 11:37 AM)
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
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Re: First attempt at agar felt like a shit show [Re: d0urd3n]
#20198606 - 06/28/14 12:00 PM (10 years, 6 months ago) |
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It gets easier with practice. You will soon learn what works and what doesn't. The main thing is that you're trying. I like to use a paper clip unfolded most of the way with the last turn left normal. Heats quick, and is sturdy enought to take the spores off the print. Keep at it and success will be yours. Good job 
Edit: always transfer at least once to ensure a clean culture. This applies for both spore and tissue inoculations.
Edited by Pastywhyte (06/28/14 12:21 PM)
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d0urd3n
Just call me "D"


Registered: 09/15/10
Posts: 5,237
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Re: First attempt at agar felt like a shit show [Re: Pastywhyte]
#20198694 - 06/28/14 12:26 PM (10 years, 6 months ago) |
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Thanks Pasty! Do you wipe down the foil on your prints? I wasn't sure if this would accomplish much.
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
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Re: First attempt at agar felt like a shit show [Re: d0urd3n]
#20198735 - 06/28/14 12:40 PM (10 years, 6 months ago) |
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Naw I don't. I usually treat prints as if they are contamed to begin with and watch carefully for first signs of germination and monitor carefully. When I make the first transfer I also look carefully for bacteria that will move out from the transfer. Usually if bacteria was present and riding along it will recover faster than the myc and you can then take action on your next transfer.
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