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Offlinesofievergais22
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Registered: 07/26/13
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Agar questions and isolating contaminated LC
    #18805868 - 09/06/13 12:01 AM (10 years, 4 months ago)

Hi everyone  i have 2 questions for y'all:

i have some agar plates ready to work with and wanted to know something, i have a SAB at the moment and i was wondering, after i cleaned my sab and added everything and then i flame sterilized my inoculation loop,do i wrap it in something or just leave it on the bottom of the SAB or on ? then spray the disinfectant...

Last question, is it worth trying to isolate a contaminated LC on agar ? or is there another way or should i just forget about it ?


Edited by sofievergais22 (09/06/13 01:50 AM)


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Offlinedeadmandave
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Re: Agar questions and isolating contaminated LC [Re: sofievergais22]
    #18806912 - 09/06/13 10:40 AM (10 years, 4 months ago)

Clean/sanitize your SAB then when everything is ready and in place, flame sterilize your inoculation loop and cool it in the agar of a the petri dish awaiting spores; the agar that just cooled the loop will help pick up spores. be sure to flame sterilize between each transfer/agar dish and dont set the sterilized instrument down or lollygag after you have flamed it because contams are still everywhere.  :biggrin:


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Offlinesofievergais22
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Registered: 07/26/13
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Last seen: 10 years, 3 months
Re: Agar questions and isolating contaminated LC [Re: deadmandave]
    #18817556 - 09/09/13 01:04 AM (10 years, 4 months ago)

Quote:

deadmandave said:
Clean/sanitize your SAB then when everything is ready and in place, flame sterilize your inoculation loop and cool it in the agar of a the petri dish awaiting spores; the agar that just cooled the loop will help pick up spores. be sure to flame sterilize between each transfer/agar dish and dont set the sterilized instrument down or lollygag after you have flamed it because contams are still everywhere.  :biggrin:




This is kind of a problem for me because if i remove my hands from the SAB then i make airflow and i have to restart all over (wait another 10 minutes) Is there no better way or am i wrong when i say i need to wait for another 10 minutes ?


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Offlinedeadmandave
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Re: Agar questions and isolating contaminated LC [Re: sofievergais22]
    #18817636 - 09/09/13 01:50 AM (10 years, 4 months ago)

dont be too afraid to take your arms in and out of the glove box; when you do move your arms make sure to have lids on petris and grain jars. only open them when you are currently doing transfers or dropping spores etc.

Make you're movements accurate and direct, soon you will gain a rhythm while doing lab work.

before you begin be sure to alcohol/sterilize the inside of the box and the surface you're working on and some people will spray the air with bleach water or lysol or something before they begin.

If we had to sterilize everything every time some air moved around we'd never get anything done; try pushing the border of what you're comfortable with when doing agar work, see what works and what doesnt - petri dishes will give fast results and no big deal if a few (dozen) contaminate :lol: .


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