transient transfection

Pamela Norton pnorton at
Tue Mar 4 13:34:40 EST 1997

In article <3314F17E.1EB7 at>, 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


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

     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

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