title: Triazole-linked DNA as a primer surrogate in the synthesis of first-strand cDNA.
abstract: 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.
authors.affiliation: Department of Chemistry, Tohoku University, Sendai, 980-8578, Japan.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Fujino
authors.forename: Ken-ichi
authors.initials: K
authors.lastname: Yasumoto
authors.forename: Naomi
authors.initials: N
authors.lastname: Yamazaki
authors.forename: Ai
authors.initials: A
authors.lastname: Hasome
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Sogawa
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Isobe
mesh_terms: D016477:Artifacts; D057930:Click Chemistry; D004247:DNA / Q000737:chemistry* / Q000235:genetics; D017931:DNA Primers / Q000138:chemical synthesis* / Q000235:genetics; D018076:DNA, Complementary / Q000032:analysis / Q000138:chemical synthesis* / Q000737:chemistry / Q000235:genetics; D005785:Gene Conversion; D043264:Phosphodiesterase I / Q000737:chemistry* / Q000235:genetics; D012333:RNA, Messenger / Q000737:chemistry* / Q000235:genetics; D012194:RNA-Directed DNA Polymerase / Q000737:chemistry / Q000235:genetics; D060327:Solid-Phase Synthesis Techniques / Q000379:methods; D014230:Triazoles / Q000737:chemistry*
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger; D014230:Triazoles; D004247:DNA; D012194:RNA-Directed DNA Polymerase; D043264:Phosphodiesterase I