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Dual phase multiplex polymerase chain reaction

United States Patent

October 7, 2008
View the Complete Patent at the US Patent & Trademark Office
Argonne National Laboratory - Visit the Technology Development and Commercialization Website
Highly specific and sensitive methods were developed for multiplex amplification of nucleic acids on supports such as microarrays. Based on a specific primer design, methods include five types of amplification that proceed in a reaction chamber simultaneously. These relate to four types of multiplex amplification of a target DNA on a solid support, directed by forward and reverse complex primers immobilized to the support and a fifth type--pseudo-monoplex polymerase chain reaction (PCR) of multiple targets in solution, directed by a single pair of unbound universal primers. The addition of the universal primers in the reaction mixture increases the yield over the traditional "bridge" amplification on a solid support by approximately ten times. Methods that provide multitarget amplification and detection of as little as 0.45-4.5.times.10.sup.-12 g (equivalent to 10.sup.2-10.sup.3 genomes) of a bacterial genomic DNA are disclosed.
Pemov; Alexander (Charlottesville, VA), Bavykin; Sergei (Darien, IL)
UChicago Argonne LLC (Chicago, IL)
10/ 794,381
March 5, 2004
The United States Government has rights in the disclosed invention pursuant to Contract No. W-31-109-ENG-38 between the U.S. Department of Energy (DOE) and the University of Chicago representing Argonne National Laboratory.