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Should I proceed with clone selection?

plasmid selection stable transfection transient transfection

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#1 BioPe

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Posted 26 December 2014 - 03:16 AM

Dear all rolleyes.gif ,

 

I transfected a cell line with a shRNA plasmid. I did a transient transfection and after 72h (which is the peak of inhibition) I could not find a decrease in the target protein.

 

In parallel, I performed a transfection for stable selection of clones. I can now see 2 or 3 clones in the plate. 

 

Is it worth keeping up with the selection of clones, since the transient transfection shows no downregulation of the target protein. Do you think the plasmid is working well?

 

Cheers and a very merry X-mas to all!

 

BioPe.



#2 bob1

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Posted Yesterday, 11:24 AM

Has the plasmid been used in the past  and worked for transient transfection? If so, it is probably worth trying again, it may be that it just didn't transfect well. It might be worth preparing a new batch of plasmid and trying that too



#3 BioPe

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Posted Yesterday, 12:35 PM

Thanks Bob,

 

I dunno if the plasmid works, it was given to me by another lab. I will give a go at the transfection again and let you know how it turns out.

Meanwhile I am still selecting de clones. I think I will check if the plasmid works in the clones too.

 

Cheers







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