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Detection of C677T mutation in methylenetetrahydrofolate reductase gene by denaturing high performance liquid chromatography
Author(s) -
Fang Nenghu,
Lin Li,
Ren Jicun,
Wu Dan
Publication year - 2004
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.364
Subject(s) - denaturing high performance liquid chromatography , methylenetetrahydrofolate reductase , chemistry , microbiology and biotechnology , dna , high performance liquid chromatography , genomic dna , gene , reductase , gel electrophoresis , chromatography , mutation , capillary electrophoresis , genetics , biochemistry , enzyme , biology , allele
In this paper, we described an assay for the detection of the C677T mutation in the methylenetetrahydrofolate reductase (MTHFR) gene using denaturing high‐performance liquid chromatography (DHPLC). The conditions for DHPLC analysis were systematically investigated based on a general HPLC instrument (Prostar VARIAN). A 225 bp DNA fragment covering the 677 site of MTHFR gene was amplied by PCR technology using the puried DNA from whole blood or whole blood as template DNA. PCR products were directly injected without the need for purication. The C677T mutation could be clearly distinguished by DHPLC technology. Our data demonstrated that DHPLC was a powerful and alternative tool for detection of genetic variants and single‐nucleotide polymorphisms to electrophoresis technology. Copyright © 2004 John Wiley & Sons, Ltd.