Colony PCR

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deedee mandark
deedee mandark's picture
Colony PCR

 I am currently attempting to establish a stable cell line using plasmid transfection system with the goal of overexpression of fully functional protein.
At the preparation of plasmid stage, I did colony PCR and successfully determined my postitive E.Coli colonies. 
I have proceeded on to transfecting my mammalian cell line (human neuroblastoma, imr 32) and the idea just hit me. 
The process of waiting: from selection to subcloning to colony selection to expansion before assessing my colony for expression  (RtPCR/W.B.) is really long.
 I just wondered if it was plausible to do the same you know, when you are sub cloning after selection to perform a 'colony PCR' at this stage. just a thought
any ideas out there?