[Analysis of temperature-modulated high-performance liquid chromatography as a tool for detection of single nucleotide polymorphism]

Abstract

Objective: To establish and evaluate temperature-modulated high-performance liquid chromatography(TmHPLC) as a rapid and efficient technique of detecting single nucleotide polymorphism(SNP).

Methods: Several TmHPLC parameters, such as the eluting gradient ranges and column temperatures used for analysing the amplicons have been studied. The size of DNA fragments suitable for analysis of SNP was studied. Also the sensitivity of the TmHPLC was evaluated.

Results: The suitable chromatographic condition of TmHPLC was found out. The elution peaks of dNTP and primers could be separated from those of polymerase chain reaction products of the genomic DNA. The heterozygotes of SNP from the sequenced DNA fragment with 178bp, 269bp and 272bp could be distinguished from the homozygotes of SNP clearly.

Conclusion: The authors have established and evaluated the TmHPLC method, which is an efficient, sensitive and rapid method for screening out SNP in the DNA fragments of larger size.