title: Rapid DNA chemical ligation for amplification of RNA and DNA signal.
abstract: Enzymatic ligation methods are useful in the diagnostic detection of DNA sequences. Here, we describe the investigation of nonenzymatic phosphorothioate--iodoacetyl DNA chemical ligation as a method for the detection and identification of RNA and DNA. The specificity of ligation on the DNA target is shown to allow the discrimination of a single point mutation with a drop in the ligation yield of up to 16.1-fold. Although enzymatic ligation has very low activity for RNA targets, this reaction is very efficient for RNA targets. The speed of the chemical ligation with an RNA target achieves a 70% yield in 5 s, which is equal to or better than that of ligase-enzyme-mediated ligation with a DNA target. The reaction also exhibits a significant level of signal amplification under thermal cycling in periods as short as 100-120 min, with the RNA or DNA target acting in a catalytic way to ligate multiple pairs of probes.
authors.affiliation: Nano Medical Engineering Laboratory, Discovery Research Institute, RIKEN, 2-1, Hirosawa, Wako-Shi, Saitama, 351-0198 Japan. h-abe@riken.jp
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Abe
authors.forename: Yuko
authors.initials: Y
authors.lastname: Kondo
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Jinmei
authors.forename: Naoko
authors.initials: N
authors.lastname: Abe
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Furukawa
authors.forename: Atsushi
authors.initials: A
authors.lastname: Uchiyama
authors.forename: Satoshi
authors.initials: S
authors.lastname: Tsuneda
authors.forename: Kyoko
authors.initials: K
authors.lastname: Aikawa
authors.forename: Isamu
authors.initials: I
authors.lastname: Matsumoto
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Ito
mesh_terms: D020137:Base Pair Mismatch; D001483:Base Sequence; D004247:DNA / Q000032:analysis* / Q000737:chemistry* / Q000235:genetics; D006358:Hot Temperature; D007461:Iodoacetates / Q000138:chemical synthesis / Q000737:chemistry; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009691:Nucleic Acid Denaturation; D009946:Organothiophosphorus Compounds / Q000138:chemical synthesis / Q000737:chemistry; D012313:RNA / Q000032:analysis* / Q000737:chemistry* / Q000235:genetics
chemical_list: D007461:Iodoacetates; D009946:Organothiophosphorus Compounds; D012313:RNA; D004247:DNA