Rapid DNA Chemical Ligation for Amplification of RNA and DNA Signal

• Published in: Bioconjugate chemistry Vol. 19; no. 1; pp. 327 - 333
• Main Authors: Abe, Hiroshi, Kondo, Yuko, Jinmei, Hiroshi, Abe, Naoko, Furukawa, Kazuhiro, Uchiyama, Atsushi, Tsuneda, Satoshi, Aikawa, Kyoko, Matsumoto, Isamu, Ito, Yoshihiro
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 01.01.2008
• Subjects: Base Pair Mismatch; Base Sequence; Biochemistry; Deoxyribonucleic acid; DNA; DNA - analysis; DNA - chemistry; DNA - genetics; Enzymes; Gene amplification; Hot Temperature; Iodoacetates - chemical synthesis; Iodoacetates - chemistry; Nucleic Acid Amplification Techniques - methods; Nucleic Acid Denaturation; Organothiophosphorus Compounds - chemical synthesis; Organothiophosphorus Compounds - chemistry; Ribonucleic acid; RNA; RNA - analysis; RNA - chemistry; RNA - genetics
• ISSN: 1043-1802; 1520-4812
• DOI: 10.1021/bc700244s

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.