DNA sequencing with chain-terminating inhibitors.

DNA sequencing with chain-terminating inhibitors.

Proceedings of the National Academy of Sciences of the United States of America (1977-12-01)

F Sanger, S Nicklen, A R Coulson

PMID271968

ABSTRACT

A new method for determining nucleotide sequences in DNA is described. It is similar to the "plus and minus" method [Sanger, F. & Coulson, A. R. (1975) J. Mol. Biol. 94, 441-448] but makes use of the 2',3'-dideoxy and arabinonucleoside analogues of the normal deoxynucleoside triphosphates, which act as specific chain-terminating inhibitors of DNA polymerase. The technique has been applied to the DNA of bacteriophage varphiX174 and is more rapid and more accurate than either the plus or the minus method.

MATERIALS

Product Number

Brand

Product Description

D7409

Sigma-Aldrich

2′,3′-Dideoxyguanosine 5′-triphosphate sodium salt, ≥90% (HPLC)

D5782

Sigma-Aldrich

2′,3′-Dideoxycytidine, ≥98% (HPLC)

D8276

Sigma-Aldrich

DNA Polymerase I, Klenow Fragment from Escherichia coli, buffered aqueous glycerol solution