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Nucleic Acids Research, Vol 25, Issue 6 1311-1312, Copyright © 1997 by Oxford University Press

ARTICLES

A DNA cycle sequencing reaction that minimizes compressions on automated fluorescent sequencers

H Yamakawa and O Ohara
Laboratory of DNA Technology, Kazusa DNA Research Institute, 1532-3 Yana, Kisarazu, Chiba 292, Japan.

We recently demonstrated that most band compressions (>70%) on DNA sequencing gels result from the presence of a single sequence motif, 5'- YGN1-2AR-3', where Y and R indicate base-pairing pyrimidine and purine residues, respectively. This finding raised the possibility that the use of 7-deaza-dATP instead of dATP in chain termination sequencing reactions would resolve most of the band compressions. Thus, we examined the effects of 7-deaza-dATP on DNA sequencing using thermostable DNA polymerases. The results indicate that the replacement of dATP with 7-deaza-dATP in conventional cycle sequencing reactions can successfully eliminate most band compressions without sacrificing sequencing performance.
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