OPTIMIZATION AND REPRODUCIBILITY OF RANDOM AMPLIFIED POLYMORPHIC DNA IN HUMAN

In this work, we have optimized random amplified polymorphic DNA (RAPD ) for the use of human DNA in altering the concentration of the reacti on components and the steps of the thermal profile in the polymerase c hain reaction. Ey using two primers in every reaction and 2.5 U Taq DN A polymerase, we found that DNA concentrations between 50 and 500 ng g ave reproducible banding patterns. The tested DNA was extracted in sev en different ways giving the same amplification results in six of them . We have also observed that reactions consisting of 35 cycles gave su fficient product yields. A slow heating/ramping from the annealing to the extension temperature increased the number of amplified bands and enhanced reproducibility. We conclude that RAPD is a robust and, under the mentioned conditions, reproducible method that could prove very u seful for scientists and physicians. (C) 1995 Academic Press, Inc.