plasmid from non viable glycerol stock

Jayakumar, R via methods%40net.bio.net (by R.Jayakumar from roswellpark.org)
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.
   
Jay


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

Hello,

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

-- 
S.G.Dalvi,
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