Hi Sunil, Glycerol stocks are stable as long as they are frozen, but tend to die if they thaw out. It is usually best to keep then at -80 and culture from scrapings off the top of the stock.
Wait for better advice on your problem, but if all else fails you could perhaps perform a miniprep on the stock, after quickly thawing, centrufuging and rinsing the cells to remove glycerol? Really though, that's an untested solution. Await a better suggestion first. Good luck, I hope you get your plasmid back! On Mar 24, 2010 1:02 PM, "Sunil Dalvi" <[email protected]> wrote: 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, _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
