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transient expression trouble

pasquale pensieri via methods%40net.bio.net (by p.pensieri87 from gmail.com)
Tue Feb 23 09:31:48 EST 2016


Hello everyone,
I would glad if someone can give me an hint about my trouble...
I would like to transfect my cell line with a plasmid GFP-Protein of interest to overexpress it. Transfection should be transient and so, after a few generation I should lose my overexpressed protein in several cells.
In any case I should select GFP population after transfection with FACS and sorting, but on these cells I would like to do mitotic spreads... So I have to put the positive cells in plate for colcemid... Could you give me an hint? If I select a population GFP positive, how can I do later to get a good number of cells for a rapid colcemid treatment in time to mantain a good number of GFP cells so as not to miss plasmid and protein, but so as I could do a good number of spreads?   
I hope I was clear...
Thank you anyone...
Pasquale


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