Sequencing directions?

Martin Hughes hughes at rkna50.riken.go.jp
Fri Feb 12 21:33:13 EST 1993


Is there any reason why it should be easier (or better) to sequence
double stranded DNA in one direction, rather than the other?

I am sequencing double stranded DNA using an automated sequencer
and the TAQ dye-primer method (ABI).  Using the M13-20 primer
I get great sequence data, but with the reverse primer, very
little at all is readable.  Since both the primers are contained in
the PRISM (pre-mixed) componant mix, I think it is unlikely
thet the difference is due to any individual pipetting or concentration
problems.  What I would like to know is if there is a "biological"
reaon behind this, or of there is a possibility that the primer kit
is at fault.

Many thanks for any replies,

Martin Hughes
Lab. Photoperception & Signal Transduction: Frontier Research:
RIKEN: Wako Shi: Saitama: JAPAN



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