salmon sperm DNA

Bernard Murray bernard at elsie.nci.nih.gov
Mon Dec 30 11:46:21 EST 1996


In article <32C2AB07.64CD at welchlink.welch.jhu.edu>, 
mathu at welchlink.welch.jhu.edu says...

[snip]
 
>As for adding this as a carrier during (mammalian?) cell tranasfections,
>again you can avoid the stuff. Supercoiled or linearized plasmid DNA
>works just fine without the carrier stuff. You have to normalize your
>transfection efficiencies using a second reporter construct anyway.

I beg to differ.  I've been trying to obtain efficient transfection of
a neuroblastoma cell line and have given it my best shot with CaPO4 and
lipofection methods.  Electroporation showed some promise and I spent
some time performing the normal optimisation for capacitance, voltage,
DNA concentration, cell density and electroporation temperature.  I saw
the biggest boost in efficiency when I included carrier DNA.  I realise
that this is not needed for all cell lines but in this case it pushed
the efficiency up to a usable level.

See Chu et al. (N.A.R. 15, 1311-1326 [1987]) for the methodology.

		Bernard

Bernard Murray, Ph.D.
bernard at elsie.nci.nih.gov (National Cancer Institute, NIH, Bethesda MD, USA)




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