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OfflineA.k.aM
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How long til you see growth of contam on agar?
    #26356705 - 12/02/19 05:51 AM (4 years, 2 months ago)

I did my first run of agar a couple days ago and took four tissue samples to clone some fruits and now I’m waiting to use the rest of the plates until I’m sure they’re clean and my sterile technique was alright.

All four clone plates look to be starting myc growth so that’s good and the unused plates still look totally clean. I need to work on eliminating condensation though a couple plates are impossible to see through from the top.

How long should I wait until declaring these plates clean?


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Offlinecurious.psychonaut
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Re: How long til you see growth of contam on agar? [Re: A.k.a]
    #26356722 - 12/02/19 06:09 AM (4 years, 2 months ago)

I had a contam appear on day 9 on a double-parafilm-wrapped dish: that's the longest I've had and I had counted it as clean by that time. Ruined a nice transfer, too. Contams can be spores and, like cube spores, they can be dry and take weeks to germinate. Out of 20 or so contam'd plates, I've only had 2 that became apparent after day 5. So, I guess, depending on your risk tolerance, wait anywhere between 1-3 weeks (or not at all)?


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OfflineA.k.aM
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Re: How long til you see growth of contam on agar? [Re: curious.psychonaut]
    #26356732 - 12/02/19 06:19 AM (4 years, 2 months ago)

Ok thanks so sounds like after 5-6 days 95% of contam will show up. I’m kinda amazed how fast the clones started growing it hasn’t even been two whole days yet.


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InvisibleSpunkyMonkey88
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Re: How long til you see growth of contam on agar? [Re: curious.psychonaut]
    #26356737 - 12/02/19 06:24 AM (4 years, 2 months ago)

Quote:

curious.psychonaut said:
I had a contam appear on day 9 on a double-parafilm-wrapped dish: that's the longest I've had and I had counted it as clean by that time. Ruined a nice transfer, too. Contams can be spores and, like cube spores, they can be dry and take weeks to germinate. Out of 20 or so contam'd plates, I've only had 2 that became apparent after day 5. So, I guess, depending on your risk tolerance, wait anywhere between 1-3 weeks (or not at all)?




Yikes... that sucks. At 3 weeks that plates gonna be damn near colonized...

I'm using glad cling wrap instead I'd parafilm u think that will be ok? Do u wrap them in the SAB?


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Re: How long til you see growth of contam on agar? [Re: SpunkyMonkey88]
    #26356753 - 12/02/19 06:40 AM (4 years, 2 months ago)

Quote:

SpunkyMonkey88 said:
I'm using glad cling wrap instead I'd parafilm u think that will be ok? Do u wrap them in the SAB?




It's in bod's TEK, so ofc it's gonna work. I've tried PVC (poor GE), LDPE (better GE), parafilm (best) & double parafilm, and noticed no difference, so I guess any wrap is at least workable (maybe in-vitro pinning is more negatively affected by poor GE though, no idea).

Edit: GLAD is LDPE


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Edited by curious.psychonaut (12/02/19 06:41 AM)


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InvisibleSpunkyMonkey88
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Re: How long til you see growth of contam on agar? [Re: curious.psychonaut]
    #26356757 - 12/02/19 06:51 AM (4 years, 2 months ago)

Quote:

curious.psychonaut said:
Quote:

SpunkyMonkey88 said:
I'm using glad cling wrap instead I'd parafilm u think that will be ok? Do u wrap them in the SAB?




It's in bod's TEK, so ofc it's gonna work. I've tried PVC (poor GE), LDPE (better GE), parafilm (best) & double parafilm, and noticed no difference, so I guess any wrap is at least workable (maybe in-vitro pinning is more negatively affected by poor GE though, no idea).

Edit: GLAD is LDPE



Good to kno, thanks. You wrap them in the SAB too?


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Offlinecurious.psychonaut
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Re: How long til you see growth of contam on agar? [Re: SpunkyMonkey88]
    #26356781 - 12/02/19 07:17 AM (4 years, 2 months ago)

I wrap them in the SAB. It's not sterile or anything (the roll is not, not to mention the gloves) but still prefer to be as clean as possible.


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OfflineA.k.aM
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Re: How long til you see growth of contam on agar? [Re: curious.psychonaut]
    #26356855 - 12/02/19 08:45 AM (4 years, 2 months ago)

I did the same also. Cling wrap done in the box. I gotta get a hack saw and chop up the roll of cling wrap it’s a pain in the ass using it normally.


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Edited by A.k.a (12/02/19 08:45 AM)


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