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can i store transformed bacteria and for how long - (Aug/28/2007 )

Hi guys, am new to this bioforum and must say i have found answers to alot of my research questions on this forum. Bravo guys!! I am newcomer to thw world of cloning, and I have a little problem, i am cloning my PCR product using the Qiagen kit using blue/white selection with kanamycin. i succesfully picked white colonies, grew them on LB with kanamycin and realised it was wknd, so i preserved them in glycerol, later on i tried to grow the bact back onto LB plates with IPTG/Xgal and kanamycin but i am got very small colonies and most of them BLUE!! What is hapening here??

-sathman-

well waht does wknd means ?????

i've stored my colonies in glycerol stock for years without pb.

-fred_33-

QUOTE (fred_33 @ Aug 28 2007, 04:00 PM)
well waht does wknd means ?????


laugh.gif wknd=weekend I presume !

You won't lose the plasmid by freezing the cells. Most likely the blue/white screening did not work first time. Did you get any blue colonies first time around or did you have a control plate that had blue colonies?

-bitesizebio guy-

yeah wknd means weekend! i had both white and blue colonies when i first plated out the bacteria, but didnt include controls without insert, so u think i should repeat the transformation step??

-sathman-

repick white and test them. Normalyl they should have kept the insert... dry.gif

-fred_33-

Thank you! will try and see what happens! biggrin.gif

-sathman-