Problem with TCA precipitation

Peter pxpst2 at vms.spam.suxs.cis.pitt.edu
Tue Mar 31 10:41:58 EST 1998


In article <3521076f.0 at newsfeed.uu.se>, "Morten Lindow"
<Morten.Lindow.5279 at student.uu.se> wrote:

> I am working with incorporation of radioactive nucleotides into the genome
> of V79 hamster fibroblasts.
> I want to determine the amount of incorporated activity.
> I have looked into protocols for DNA precipitation with TCA, but they all
> seem to be on purified DNA.
> Does anyone know how to apply this method (or an alternative) to whole
> cell-systems?
> A quick and safe lysing maybe?

We do in cell culture.
We remove culture media then apply a 1.5 ml cold 5% TCA to cover the
entire well.
We allow it to sit in the cold room for 30 minutes then we rmove TCA and
"dip" wash the plates.  Allow them to dry completely then solubulize DNA
with .3M NaOH and count.
Peter

-- 
"Don't you eat that yellow snow
            watch out where the Huskies go"    FZ

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