METHODS FOR SIMULTANEOUS AMPLIFICATION OF TARGET LOCI

摘要

The invention provides methods for simultaneously amplifying multiple nucleic acid regions of interest in one reaction volume as well as methods for selecting a library of primers for use in such amplification methods. The invention also provides library of primers with desirable characteristics, such as minimal formation of amplified primer dimers or other non-target amplicons.

主权项

1. A method of amplifying target loci in a nucleic acid sample, the method comprising:
(a) contacting a sample comprising target human loci with a library of at least 50 non-immobilized, non-identical primers that simultaneously hybridize to at least 50 non-identical target human loci to produce a reaction mixture; wherein the primers do not include molecular inversion probes (MIPs); (b) subjecting the reaction mixture to primer extension reaction conditions to produce amplified products comprising target amplicons; wherein the annealing temperature for the reaction conditions is greater than a melting temperature of the at least 50 non-identical primers; wherein the length of the annealing step of the reaction conditions is greater than 5 minutes; and wherein at least 50 non-identical target human loci are simultaneously amplified; and (c) sequencing the amplified products.