1. Academic Validation
  2. Improvements in the chain-termination method of DNA sequencing through the use of 7-deaza-2'-deoxyadenosine

Improvements in the chain-termination method of DNA sequencing through the use of 7-deaza-2'-deoxyadenosine

  • DNA Seq. 1991;1(4):233-9. doi: 10.3109/10425179109020778.
M A Jensen 1 R J Zagursky G L Trainor A J Cocuzza A Lee E Y Chen
Affiliations

Affiliation

  • 1 Central Research and Development Department, E.I. du Pont de Nemours & Company (Inc.), Wilmington, DE 19898.
Abstract

Significant improvements in the quality of DNA sequencing data have been shown when deoxyadenosine triphosphate (dATP) is replaced by 7-deaza-2'-deoxyadenosine triphosphate (c7dATP). The use of c7dATP in conjunction with 7-deaza-2'-deoxyguanosine triphosphate (c7dGTP) further decreases anomalies in electrophoretic mobility which are caused by compressions involving G and/or A residues. This effect is observed for both isotope-based and fluorescence-based sequencing approaches. Replacing dATP with c7dATP also results in a higher degree of uniformity in the frequency of chain termination reactions, when such terminations involve the incorporation of fluorescence-labeled dideoxynucleotides by T7 polymerase. These improvements in the gel-resolution and distribution of chain-terminated DNA products result in higher accuracy in both manual and automated base assignment.

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