Premium
Triazole‐Linked DNA as a Primer Surrogate in the Synthesis of First‐Strand cDNA
Author(s) -
Fujino Tomoko,
Yasumoto Kenichi,
Yamazaki Naomi,
Hasome Ai,
Sogawa Kazuhiro,
Isobe Hiroyuki
Publication year - 2011
Publication title -
chemistry – an asian journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.18
H-Index - 106
eISSN - 1861-471X
pISSN - 1861-4728
DOI - 10.1002/asia.201100712
Subject(s) - complementary dna , reverse transcriptase , primer (cosmetics) , oligonucleotide , exonuclease , biology , dna , microbiology and biotechnology , transcription (linguistics) , polymerase chain reaction , computational biology , chemistry , genetics , polymerase , gene , linguistics , philosophy , organic chemistry
A phosphate‐eliminated nonnatural oligonucleotide serves as a primer surrogate in reverse transcription reaction of mRNA. Despite of the nonnatural triazole linkages in the surrogate, the reverse transcriptase effectively elongated cDNA sequences on the 3′‐downstream of the primer by transcription of the complementary sequence of mRNA. A structure–activity comparison with the reference natural oligonucleotides shows the superior priming activity of the surrogate containing triazole‐linkages. The nonnatural linkages also protect the transcribed cDNA from digestion reactions with 5′‐exonuclease and enable us to remove noise transcripts of unknown origins.