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In addition, our assay allows the design for signal branching from a string of straight-line starters of 2 5 proteins or more in the relay.
CP developed the assay, designed the study, helped perform the experiments, analyzed the data, and drafted the manuscript.
MRH helped with the assay design and the draft of the manuscript.
PCR primers were designed using Primer3 software located at the "Assay Design Center" of the Roche web-site.
The other 12 SNPs' genotyping was carried out with iPLEX assays (Sequenom, San Diego, CA, USA), which were designed by the Assay Design Suite 1.0 online software.
These high-quality SNP were designated for inclusion in the assay design for use in parentage validation.
The assay design and conditions were based on the allelic discrimination protocol from Applied Biosystems.
The assay design includes an amplification control DNA template for the recognition of PCR-inhibitors.
Of the 3,230 SNPs, 3% failed as a result of the assay design.
NEM was responsible for the assay design, measurements of NGAL, statistical analyses and for writing the paper.
The assay design principles in this paper are also relevant to the HRM screening of other germline mutations.
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