title: Improvements in the chain-termination method of DNA sequencing through the use of 7-deaza-2'-deoxyadenosine.
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.
authors.affiliation: Central Research and Development Department, E.I. du Pont de Nemours & Company (Inc.), Wilmington, DE 19898.
authors.forename: M A
authors.initials: MA
authors.lastname: Jensen
authors.forename: R J
authors.initials: RJ
authors.lastname: Zagursky
authors.forename: G L
authors.initials: GL
authors.lastname: Trainor
authors.forename: A J
authors.initials: AJ
authors.lastname: Cocuzza
authors.forename: A
authors.initials: A
authors.lastname: Lee
authors.forename: E Y
authors.initials: EY
authors.lastname: Chen
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D005821:Genetic Techniques; D008969:Molecular Sequence Data; D013728:Terminator Regions, Genetic; D014372:Tubercidin / Q000031:analogs & derivatives* / Q000737:chemistry
chemical_list: C053222:2'-deoxytubercidin; D000255:Adenosine Triphosphate; D004247:DNA; D014372:Tubercidin