plasmid from non viable glycerol stock

Jayakumar, R via (by R.Jayakumar from
Wed Mar 24 08:15:43 EST 2010

What percentage glycerol did you use nad was this frozen at -80C?  I use 40% glycerol to freeze and it lasts for years and years with good viability. Moreover, it is important that you never thaw the glycerol stock inbetween uses.  That reduces viability too.  I doubt whether you can retrieve the plasmid DNA intact from a lysed bacterial stock.  But it would not hurt to do a plasmid prep on the glycerol stock and see whether anything has survived DNAses.

-----Original Message-----
From: methods-bounces from [mailto:methods-bounces from] On Behalf Of Sunil Dalvi
Sent: Wednesday, March 24, 2010 1:51 AM
To: methods from
Subject: plasmid from non viable glycerol stock


I am working with Agrobacterium transformation of sugarcane.I prepared the
glycerol stock of plasmid transformed in E.Coli DH5 alpha and EHA 105. After
some months those are non reviving. What may be the problem? Is there any
method to recover plasmid from glycerol stocks.

Please help me

Is there anybody who can help me

Methods mailing list
Methods from

This email message may contain legally privileged and/or confidential information.  If you are not the intended recipient(s), or the employee or agent responsible for the delivery of this message to the intended recipient(s), you are hereby notified that any disclosure, copying, distribution, or use of this email message is prohibited.  If you have received this message in error, please notify the sender immediately by e-mail and delete this email message from your computer. Thank you.

More information about the Methods mailing list