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transient transfection

Wed Feb 26 21:30:36 EST 1997

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

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