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

F. Fernandez-Rachubinski Department of Pathology, McMaster University, Hamilton, Ontario, Canada, L8N 3Z5

W. W. Murray Department of Biochemistry, McMaster University, Hamilton, Ontario, Canada, L8N 3Z5

M. A. Blajchman Department of Pathology, McMaster University, Hamilton, Ontario, Canada, L8N 3Z5; Canadian Red Cross Society Blood Transfusion Service, Hamilton, Ontario, Canada, L8N 3Z5

R. A. Rachubinski Department of Biochemistry, McMaster University, Hamilton, Ontario, Canada, L8N 3Z5

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 ONA by the dideox-yribonucleotide chain termination method.

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

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

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