R to DK: How to efficiently transfect U937 cells with a plasmid vectorexpressing shRNA

Prof. Piero Sestili via methods%40net.bio.net (by piero.sestili from uniurb.it)
Wed Aug 8 08:37:13 EST 2007


Thanks a lot DK,  

we have something like this guide but there's nothing "amazing"....so
yes, please send your one to the public address or, if you prefer to
piero.sestili from uniurb.it. Hi and thanks again

Piero

Prof. Piero Sestili
Istituto di Farmacologia e Farmacognosia e
Centro di Ricerca sull'Attività Motoria
Università degli Studi di Urbino "Carlo Bo"
Via "I Maggetti" 26
61029 URBINO (PU)
Tel. 0722 303414; 0722 305524 
Fax 0722 303401


-----Messaggio originale-----
Da: methods-bounces from oat.bio.indiana.edu
[mailto:methods-bounces from oat.bio.indiana.edu] Per conto di DK
Inviato: martedì 7 agosto 2007 23.51
A: methods from magpie.bio.indiana.edu
Oggetto: Re: How to efficiently transfect U937 cells with a plasmid
vectorexpressing shRNA

In article <mailman.546.1186502638.11350.methods from net.bio.net>, "Prof.
Piero Sestili" <piero.sestili from uniurb.it> wrote:
>We are trying to transfect U937 cells with a plasmid vector expressing 
>shRNA. However we are obtaining a very low transfection efficiency 
>using either electroporation (which is not the best solution since we 
>need that transfected cells retain their viability for some weeks) or 
>lipofectin.  Is there anybody who knows some nice alternative method or

>trick to increase the efficiency?  thanks a lot,

The highest efficiency is practically always with electroporation. The
trick is that it needs to be properly optimized. If you would 
you be willing to go this route, I can post an "electroporation 
optimization guide" that I've used to transfect all kinds of cells and
help many people over many years. 

DK
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