Trouble with CaPO4 transfections

Helge Steen mbxhhs at mbn1.biochem.nottingham.ac.uk
Thu Sep 26 13:33:59 EST 1996


Hi,

We've been using the CaPO4 method to transfect mammalian cells (eg NIH
3T3) successfully for years.
After a recent lab move we are now having problems because the apparent
transfection efficiency is gradually decreasing. A recent test using
X-gal staining revealed that the transfection efficiency is approx. 1%.
This is less than 1/10th the efficiency we had before.

The precipitate we see is comparable to that we saw before the move and
neither of our assays (luciferase & beta-gal) have changed.

If anyone has experienced a similar problem we would love to know what
the problem was with your system. 
Any advice gratfully recieved,

Thanks, Helge



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