Abstract
'MegaPlex PCR' is a robust technology for highly multiplexed amplification of specific DNA sequences. It uses target-specific pairs of PCR primers that are physically separated by surface immobilization. Initial surface-based amplification cycles are then coupled to efficient solution-phase PCR using one common primer pair. We demonstrate this method by co-amplifying and genotyping 75 unselected human single-nucleotide polymorphism (SNP) loci.
Highlights
During the development of MegaPlex PCR we evaluated a range of targets, input DNAs and reaction conditions
Using the microbead support we found MegaPlex PCR to be effective with as little as 200 ng of human genomic DNA, and in various 15-plex reactions it recovered many different target sequences of up to at least 500 bp in size, with little bias against larger amplicons (Supplementary Fig. 1 online)
The protocol used microbeads adhered to the walls of microtiter plate wells as the solid surface, and genomic DNA samples enriched for the MP75 and the MP50 sets of targets
Summary
During the development of MegaPlex PCR we evaluated a range of targets, input DNAs and reaction conditions. Using the microbead support we found MegaPlex PCR to be effective with as little as 200 ng of human genomic DNA, and in various 15-plex reactions it recovered many different target sequences of up to at least 500 bp in size, with little bias against larger amplicons (Supplementary Fig. 1 online).
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