An allylic/acyclic adenosine nucleoside triphosphate for termination of DNA synthesis by DNA template-dependent polymerases

An allylic adenosine triphosphate analog (AATP) was tested as a substrate f or commercially available DNA polymerases. All but one of the enzymes assay ed incorporated AATP opposite thymidine (T) with concomitant termination of the elongation reaction. A concentration of only 1 mu M was sufficient for complete termination of the polymerization reaction for a short template m ediated by Ampli Tag DNA polymerase FS (Taq FS). This result suggests that AATP could be used as a 2',3'-dideoxyadenosine-5'-triphosphate (ddA) surrog ate. Kinetics of incorporation revealed that AATP was 48 times less efficie ntly incorporated than ddA. Furthermore, AATP was used in dye-primer sequen cing as a substitute for ddA.