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??
well waht does wknd means ?????
i've stored my colonies in glycerol stock for years without pb.
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?
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??
repick white and test them. Normalyl they should have kept the insert...
Thank you! will try and see what happens!