Clone selection for scrambled shRNA transfected cells ? - (Sep/16/2013 )

Hi all !

I'm starting to work on gene silencing and I have little in the subject.

I did shRNA transfections en a cell line using plasmids and I am now in the stage of clonal selection, to establish monoclonal populations for stablishment of a stable transfected cell line.

My question is: what is the relevance of establishing a monoclonal cell line with the control plasmid (scrambled seq) ?.... or is it sufficient for future functional assays to use the polyclonal populations as negative control effects.

I hope the question is clear ... I´ll be very gratefut to get any information or reading material on this topic.

Thanks!

-anahifranchi-

You also need to establish a monoclonal population of cells which carry the plasmids expressing the shRNA (control) sequence in the exactly same way as you create the shRNA of interest cell line.

-pcrman-

Thank you!

-anahifranchi-