DNA polymerases free of bacterial (DNA) contamination
rfwhittier at hotmail.com
Sun Mar 9 21:45:19 EST 2003
Duncan Clark wrote/schrieb
>Historians believe that in newspost
><e05b55b0.0303050718.113abe56 at posting.google.com> on Wed, 5 Mar 2003,
>Mashounya <oldhorse at mail.ru> penned the following literary masterpiece:
>>I routinely work with universal bacterial primers specific to 16S rRNA
>>gene and I need Taq polymerase free of bacterial DNA. I nave no time and
>>money to check different commercially available polymerases. If anyone
>>working in this area has chosen a good supplier (really high quality, not
>>50/50), please, inform me. I've tried Promega PCR Master Mix - one lot was
>>suitable, another - not.
>I do not think you will find any.
>Check out using psoralen and UV
>Nucleic Acids Res 1990 Nov 25;18(22):6739 Use of psoralen as extinguisher
>of contaminated DNA in PCR.
>Jinno Y, Yoshiura K, Niikawa N.
Here's another simpler but less elegant suggestion.
If the archaebacterial sequence differs sufficiently
from the eubacterial sequence, just purchase *native*
Taq polymerase instead. It is still contaminated. but
the contamination is different. I used this solution
when contaminating common vector sequences were
wreaking havoc on my ability to recover flanking
sequence tags from mouse insertions by TAIL-PCR. The
native Taq did not have the plasmid sequences.
You should be able to find the native Taq polymerase
in the Amersham Biosciences catalog, and while I have
not checked, I would guess that other outfits also
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