Feliciello_2004_Nutr.Metab.Cardiovasc.Dis_14_205

We developed a rapid procedure to analyse simultaneously two different DNA polymorphisms of the human LPL gene by PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism). The method involves PCR amplification of the gene fragments encompassing two polymorphic sites, direct digestion in the same PCR-tube of the amplification mixture with two restriction enzymes, and the analysis of the resulting DNA fragments by gel electrophoresis. In 422 participants of the 1994 follow-up examination of the Olivetti Heart Study, a total of eight common LPL gene polymorphisms have been analysed in pairs by this procedure: -93 T/G and D9N; V108V and T361T; N291S and PvuII; HindIII and S447X. Two of these polymorphisms (V108V and T361T) were analysed for the first time. This method is suitable for the routine analysis of clinical samples of varying DNA content and practically halves the times and costs of screening for these LPL polymorphisms.