Genotyping single nucleotide polymorphisms by primer extension and high performance liquid chromatography

We have investigated the possibility of genotyping single nucleotide polymo rphisms (SNPs) by primer extension and high performance liquid chromatograp hy (HPLC). Using three polymorphisms of current interest to our group (an A /G polymorphism in the proneurotensin gene and A/G and T/C polymorphisms in the 5HT2a receptor gene), we show that robust signal is obtained using thi s simple analytic method which has the added advantages that sample loading and analysis are essentially automated, analytic time is brief, and no fur ther purification step after primer extension is required. We also show tha t all stages of the HPLC-primer extension genotyping can be multiplexed whi ch, together with automation, suggests that this system may be suitable for linkage studies based upon emerging SNP maps.