earticle

영어

Single nucleotide difference discrimination is important for field of genotyping including Single Nucleotide Polymorphism (SNP). Several technical approaches were applied for discriminating single nucleotide difference discrimination, ligation based technologies were broadly used which use DNA ligase and target specific probe set. Ligation based technology genotype the single nucleotide with ligation efficiency difference between perfectly matched probe and mismatched probe. If 3’' end of left probe oligo (LPO) hybridize perfectly with target site, LPO ligated with 5’' end of right probe oligo nucleotide (RPO). But in the case of mismatch between 3’' end of LPO and target site, ligation between LPO and RPO is still occurred in some case because of high reaction rate of DNA ligase, so it does not gives accurate information about genomic information of target site. We used commercially available alternative nucleobase such as 5’'-nitroindole, iso dC, iso dG in 5’' end of RPO to solve the problem by decreasing reaction rate of DNA ligase. By this approach, various matches and mismatch between 3’' end of LPO and target site was discriminated.