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Nucleic Acids Symposium Series 2003 3(1):37-38; doi:10.1093/nass/3.1.37
© 2003 by Oxford University Press
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Simultaneous incorporation of three different modified nucleotides during PCR

Masayasu Kuwahara, Shin-ichi Hososhima, Yumi Takahata, Rina Kitagata, Atsushi Shoji, Kazuo Hanawa, Akiko N. Ozaki, Hiroaki Ozaki and Hiroaki Sawai

Department of Applied Chemistry, Gunma University, Kiryu, Gunma 376-8515, Japan

Modified analogs of 2'-deoxycytidine triphosphates bearing (6-aminohexyl)carbamoylmethyl or 7-amino-2,5-dioxaheptyl Linker at a C5 position were designed and synthesized. Both analogs were found to be good substrates for Vent(exo-) DNA polymerase during PCR, resulting in the corresponding full-length modified DNAs, respectively. Moreover, we have demonstrated simultaneous incorporation of three different modified nucleotides into a DNA strand by PCR using triphosphates of 5-(3-aminopropynyl)dUTP, 5-[(6-aminohexyl)carbamoylmethyl]dCTP and 2-amino-dATP (dDTP) or N6-methyl-dATP in place of the natural nucleoside triphosphates TTP, dCTP and dATP.


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