Trouble with CaPO4 transfections

JAne janeh at ihug.co.nz
Tue Oct 1 15:00:23 EST 1996


In article ,  says...
>
>Helge Steen wrote:
>>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 luciferase assay has not changed, perhaps you should try a 
new supply
>of X-gal.  Your transfection efficiency has probably not 
changed.
>
>Krista Evans
>kevans at lifetech.com


Helge,
we had exactly the same after moving to new labs. Turned out to 
be the water quality, believe it or not.
KLAUS




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