Incorporation of 7-deaza dGTP during the amplification step in the polymerase chain reaction procedure improves subsequent DNA sequencing

DNA Seq. 1990;1(2):137-40. doi: 10.3109/10425179009016041.

Abstract

An improved method for sequencing human genomic DNA amplified by the polymerase chain reaction (PCR) procedure is described. The portion of the genome investigated is the 383 nucleotide-long exon 2 of the human antithrombin III gene. Incorporation of the analogue of dGTP, 7-deaza-2'-deoxyguanosine-5'-triphosphate, during the amplification of exon 2 by PCR allowed for the elimination of recurrent artifacts obtained during sequencing of the amplified DNA by the dideoxyribonucleotide chain termination method.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antithrombin III / genetics
  • Base Sequence
  • Blotting, Southern
  • DNA
  • Guanine / analogs & derivatives*
  • Guanine / chemistry
  • Humans
  • Molecular Sequence Data
  • Polymerase Chain Reaction / methods*

Substances

  • Guanine
  • Antithrombin III
  • DNA
  • 7-deazaguanine