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3′‐Amino‐Modified Nucleotides Useful as Potent Chain Terminators for current DNA sequencing methods
Author(s) -
Herrlein Mathias K.,
Konrad Renate E.,
Engels Joachim W.,
Holietz Torsten,
Cech Dieter
Publication year - 1994
Publication title -
helvetica chimica acta
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.74
H-Index - 82
eISSN - 1522-2675
pISSN - 0018-019X
DOI - 10.1002/hlca.19940770222
Subject(s) - chemistry , nucleotide , dna , dna sequencing , computational biology , combinatorial chemistry , biochemistry , gene , biology
We describe the synthesis of modified nucleoside triphosphates of the four DNA bases containing a 3′‐amino group which were prepared from the corresponding 3′‐azido derivatives. Introduction of the triphosphate and subsequent reduction of the N 3 to the NH 2 group led directly to the target molecules 6a–d . Furthermore, 3′‐amino‐2′,3′‐dideoxynucleoside 5′‐triphosphates proved to be potent inhibitors of the enzymatic synthesis of DNA catalyzed by the standard sequencing enzymes T7 DNA polymerase, sequenase version 2.0, Thermus aquaticus DNA polymerase, and Thermus thermophilus DNA polymerase. Both radioactive and fluorescent sequencing methods were applied successfully to the 3′‐amino‐modified terminators. Investigations in view of using these chain terminators according to Sanger's sequencing method for fluorescence labeling were done.