sequencing problems w/ ALF II automated sequencer

Gizbab gizbab at aol.com
Mon Mar 31 21:20:18 EST 1997


Hi!

I wonder if anyone has run into this problem.... I have had some problem
with the ALF II automated sequencing machine. Namely, when I work with
cell lines, the sequences come up fine, yet when I use tissues extracted
from paraffin, and sequence the stuff for p53 mutations (homo and
heterozygous), I get some strange results, if any! It got to the point
htat I simply took some of the cell line DNA, amplified it for one of the
p53 exons, and cycle sequenced it all in one reaction tube, and then ran
the product in all lines of the machine and actually found different
results in most of the cases! I really have little problem sequencing the
conventional way (radioactive) and get great results that way. Any help
for user of the ALF II will be very much appreciated.

Ron Przygodzki, MD
gizbab at aol.com



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