transient transfection

Pamela Norton pnorton at lac.jci.tju.edu
Tue Mar 4 13:34:40 EST 1997


In article <3314F17E.1EB7 at mcleod.net>, l.peterson at MCLEOD.NET (PETERSON
LAWRENCE) wrote:

> I need help trying to figure out why my transfections aren't working 
> very well.  I am using NIH 3T3 cells and Lipofectin.  I have tried 
> various ratios of Lipofectin:DNA, various times of incubation, with 
> Optimem or with other serum free medium.  I am currently just 
> transfecting a reporter vector (pSVB-Gal) and then using an X-Gal assay 
> to identify transfected cells.  Way less than 1% of the cells are 
> getting transfected.  Everyone says these cells are so easy to transfect 
> and I should be able to do it blind folded.  What am I doing wrong? 
> 
> I start with cells that are 60 to 80% confluent and then do my assay 
> either 24 or 48 hours later.
> 
> Any insights would be greatly appreciated. 
> 
> Rebecca Slayton
> email: rebecca-slayton at uiowa.edu

Hi, 

     We also failed to get good transfection of NIH 3T3 cells with
Lipofectin. Try Lipofectamine, it works _much_ better for 3T3s. (No
affiliation....)

     Mail me if you would like more information.

     Good luck,

          Pam Norton

-- 
Pamela A. Norton, Ph.D.          Associate Professor of Medicine
Thomas Jefferson University
Philadelphia, PA 19107           p_norton at lac.jci.tju.edu



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