PCR with radioactive nucleotides

M K Bennett pamkb at mail.bris.ac.uk
Wed Nov 16 10:58:39 EST 1994


ROSS at SCI.WFEB.EDU wrote:

: We have been doing PCR with S35 nucleotides and have just found that
: the PCR block is heavily contaminated.  The manufacturer of the PCR
: says that S35 can go through the walls of the tube.  This is a little
: hard for me to believe.  I wonder if some tubes do not seal properly.
: In any case,  have other people had this problem and is there any
: way to avoid this type of contamination??

This is a well known finding and is due to the volatility of the 35S
nucleotides.  They escape from the tubes through the lids because they
dont seal that well.

As to avoiding it use 33P rather than 35S.



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