Study of the base discrimination ability of DNA and 2′- O-methylated RNA oligomers containing 2-thiouracil bases towards complementary RNA or DNA strands and their application to single base mismatch detection

Abstract

Precise detection of target DNA and RNA sequences using chemically modified oligonucleotides is of crucial importance in gene analysis and gene silence. The hybridisation and base discrimination abilities of oligonucleotides containing 2′- O-methyl-2-thiouridine (s 2Um) in homo- and hetero-duplexes composed of DNA and RNA strands have been studied in detail. When s 2Um was incorporated into RNA or DNA strands, the hybridisation and base discrimination abilities of the modified RNA or DNA oligomers towards the complementary RNA strands were superior to those of the corresponding unmodified oligomers. On the other hand, their base discrimination abilities towards complementary DNA strands were almost the same as those of the unmodified ones. The base discrimination abilities of 2-thiouracil base-containing oligonucleotide probes on slide glass plates were also studied. These modified probes exhibited efficient detection of mismatched base pairing.