Improvements in the chain-termination method of DNA sequencing through the use of 7-deaza-2'-deoxyadenosine.
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.
Jensen
M A
MA
Central Research and Development Department, E.I. du Pont de Nemours & Company (Inc.), Wilmington, DE 19898.
Zagursky
R J
RJ
Trainor
G L
GL
Cocuzza
A J
AJ
Lee
A
A
Chen
E Y
EY
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
C053222:2'-deoxytubercidin; D000255:Adenosine Triphosphate; D004247:DNA; D014372:Tubercidin