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
> 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
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.
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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