SIMULTANEOUS-DETERMINATION OF METHYLENETETRAHYDROFOLATE REDUCTASE C677T AND FACTOR-V G1691A GENOTYPES BY MUTAGENICALLY SEPARATED PCR AND MULTIPLE-INJECTION CAPILLARY ELECTROPHORESIS
A. Ulvik et al., SIMULTANEOUS-DETERMINATION OF METHYLENETETRAHYDROFOLATE REDUCTASE C677T AND FACTOR-V G1691A GENOTYPES BY MUTAGENICALLY SEPARATED PCR AND MULTIPLE-INJECTION CAPILLARY ELECTROPHORESIS, Clinical chemistry, 44(2), 1998, pp. 264-269
The C677T mutation of the methylenetetrahydrofolate reductase gene and
the G1691A (Leiden) mutation of the factor V gene are established ris
k factors for thromboembolic disease. We here present an assay for the
simultaneous genotyping of these common genetic variants. The assay i
nvolves a strategy based an multiplex mutagenically separated PCR perf
ormed in a single tube containing six primers. Separation of the resul
ting four PCR products (197, 207, 233, and 246 bp) was performed by ca
pillary electrophoresis coupled with laser-induced fluorescence detect
ion. The time for the automated electrophoresis was reduced to 2.5 min
per sample by performing the capillary electrophoresis analysis in a
multiple-injection mode.