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colonies on a plate


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2 replies to this topic

#1 claritylight

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Posted 21 May 2009 - 05:36 AM

I streaked out some colonies on some LB+ Amp plates back in December 2008 for DNA sequencing. I picked out the colonies for my experiment and then stored the plates in the 4*C since then. I want to repeat the experiment again now using these plates.

Is this a good way for long term storage of these plates?

#2 phage434

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Posted 21 May 2009 - 05:55 AM

The cells on those plates are almost certainly dead. For long term storage, grow cultures of your cells in LB or another rich medium, add sterile 80% glycerol solution to give a final 15% glycerol concentration in your medium, and freeze at -80C for storage indefinitely. If you are desperate to recover a plasmid from your (now dead) cells on the plate, it might be possible to prep DNA and transform living cells from your colonies, but I would not count on it. Storing plasmid DNA is also an effective means of long term storage.

#3 Vini

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Posted 22 May 2009 - 01:39 AM

The cells on those plates are almost certainly dead. For long term storage, grow cultures of your cells in LB or another rich medium, add sterile 80% glycerol solution to give a final 15% glycerol concentration in your medium, and freeze at -80C for storage indefinitely. If you are desperate to recover a plasmid from your (now dead) cells on the plate, it might be possible to prep DNA and transform living cells from your colonies, but I would not count on it. Storing plasmid DNA is also an effective means of long term storage.



yeah, i agree with that. i myself have tried to revive colonies from a month old plate, it did not work....i wouldn't rely on such an old plate.




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