Controlling performance of complex polymerase chain reaction amplification, useful e.g. for genotyping, using a set of many specific primers and non-specific counter-strand primers

摘要

Controllable performance method of complex polymerase chain reaction (PCR) amplifications. First, PCR is carried out with at least 50 different primers (P1) of one type, complementary to one strand of sample DNA, and with a primer (or library of primers) of a second type (P2) complementary to the other strand of the DNA, with P2 carrying a marker (M1). Amplicons are hybridized either to an array of oligonucleotides (ON) that hybridize to the primer used for the first step, or to its complement, or to an array of ON complementary to the primers used in PCR, and then the lengths of amplicons bound to the array are determined using a second marker (M2), different from M1, that is correlated with the length of the relevant DNA fragments. Signals from M1 and M2 are quantified at relevant positions in the ON array.