A process for the detection of cytochrome P450 2C9 gene variants

Abstract

This invention relates to a process for detection of cytochrome P450 2C9 gene variants in the human genomic DNA in PCR tubes containing 0.74 U of Taq polymerase, 2 mM MgCl2, 200 mM each deoxynucleotidetriphosphate, 0.5 pmol of CYP2C9*2wild-type reverse primer (Primer 1), 1.5 pmol of CYP2C9*2 mutant reverse primer (Primer 2), 3 pmol of CYP2C9*2 common forward primer (Primer 3),1 pmo1 of CYP2C9*3 wild-type forward primer (Primer 4), 2 pmol of CYP2C9*3 mutant forward primer (Primer 5), and 3 pmol of CYP2C9*3 common reverse primer (Primer 6), and ~2 ng of DNA and performing amplifications in an Eppendorf Thermal Cycle of 5-min denaturation period at 95°C followed by 35 cycles of 95 °C for 45 s, 57°C for 45 s, and 72°C for 45 s, whereby a final extension step of 10 min at 72°C completes the reaction and using (a) Primer sequence land 3 for the determination of *1 *1 genotype (b) Primer sequence 1, 2 and 3 for the determination of * 1 *2 genotype (c) Primer sequence 2 and 3 for the determination of *2*2 genotype (d) Primer sequence 4 and 6 for the determination of *1 *1 genotype (e) Primer sequence 4, 5 and 6 for the determination of *1 *3 genotype and (f) Primer sequence 5 and 6 for the determination of *3*3 genotype.