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Clone selection for scrambled shRNA transfected cells ?

shRNA negative control

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#1 anahifranchi

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Posted 16 September 2013 - 09:22 AM

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!


#2 pcrman

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Posted 16 September 2013 - 10:36 AM

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. 



#3 anahifranchi

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Posted 17 September 2013 - 02:49 PM

Thank you!






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