Jump to content

  • Log in with Facebook Log in with Twitter Log In with Google      Sign In   
  • Create Account

- - - - -

colonies on a plate


  • Please log in to reply
2 replies to this topic

#1 claritylight

claritylight

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 104 posts
0
Neutral

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

phage434

    Veteran

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 1,846 posts
142
Excellent

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

Vini

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 178 posts
1
Neutral

Posted 22 May 2009 - 01:39 AM

View Postphage434, on May 21 2009, 06:55 AM, said:

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.




Home - About - Terms of Service - Privacy - Contact Us

©1999-2012 Protocol Online, All rights reserved.