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Enzymatic synthesis of modified DNA by PCR
M M Masud1, A Ozaki-Nakamura, F Satou
1Department of Applied Chemistry, Faculty of Engineering, Gunma University, Kiryu, Gunma 376-8515, Japan.
Nucleic Acids Research. Supplement (2001)
|July 3, 2003
Abstract:
We synthesized various 5'-triphosphates of C5-substituted 2'-deoxyuridine derivatives bearing methylene linker at C5-alpha position. We examined whether the C5-substituted 2'-deoxyuridine 5'-triphosphates (dUTP) can work as a substrate for the modified DNA synthesis by PCR. We found that only KOD dash DNA polymerase, a thermostable DNA polymerase from extremely thermophilic archaeum, accepted the modified substrates in place of TTP for PCR forming the corresponding modified DNAs. On the other hand, no other DNA polymerase could accept these TTP analogues.