Transfection grade plasmid DNA

Sat Sep 3 10:32:29 EST 1994

Hello netters,

Our lab is preparing to do some eukaryotic

cell transfections, an area that we don't

have any experience in.  I have heard from 

colleagues that getting very pure plasmid 

preparations (2 X banded in a CsCl gradient)

is critical for high efficiency and 

reproducibility.  My questions are:

1.	Is this true?  What is the best

method for purifying plasmids for 

transfection?  Do commercially-available

column methods work at all?  If so, 

which ones are best?

2.	Have there been any studies

or publications on transfection efficiencies

using different plasmid purification methods?

If so, what are the references?  It all seems

a little like voodoo and heresay to me.  I would

like to see the science behind this issue.

3.	What is the cause of low transfection

 efficiencies in certain plasmid preps?  Is protein

 or nucleic acid contamination important?  Do 

endotoxins have anything to do with it?  

Thanks in advance for your help.

Rick (in Indiana)


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