IUBio

Trichloroacetate and nucleic acids

Ned Mantei mantei at cell.biol.ethz.ch
Tue Mar 23 04:23:49 EST 1999


In article
<Pine.SUN.3.92.990323143828.5250A-100000 at molecule.bio.uts.edu.au>,
michelle at MOLECULE.BIO.UTS.EDU.AU (Michelle Gleeson) wrote:

> Hi Emir,
> 
> Chloroform might work for your purposes - it should kill the bacteria very
> quickly, and won't damage the DNA.  However, the catch is that the
> culture vessels, plates etc will need to be glass not plastic. Check out
> some bacteriophage isolation methods for the details.

Chloroform will kill bacteria within about 30 seconds. See:

Nature 1976 Jun 3;261(5559):428-9 
Reduction of possible hazards in the preparation of recombinant plasmid DNA.
Weissmann C, Boll W

However, the cells mostly don't lyse, and it is not at all clear that
nucleases would be inhibited. Incidentally, polypropylene plastic will
resist chloroform for quite awhile.
Trichloroacetic acid, as suggested by the original poster, will cause
depurination of DNA and RNA,and is therefore not suitable.
It is possible to filter off the bacteria within a few seconds. It should
then be possible to very rapidly freeze the bacteria on the filter (dry
ice or liquid nitrogen), thus stopping everything.

-- 
Ned Mantei
Dept. of Cell Biology, Swiss Federal Institute of Technology
CH-8093 Zurich, Switzerland



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