Company: Pioneer Hi-Bred
Liaison(s): Wen-Chy Chu
This Team Masters Project utilized the results generated from the 2004-2005 academic year TMP with Pioneer Hi-Bred Molecular Markers Group and focused on the development of a high-throughput SNP detection assay for maize genotyping. A novel nucleic acid amplification assay developed at KGI, the ProxAR (Proximity Amplification Reaction) scheme is based on a tri-body hybridization coupled with a molecular beacon fluorescent readout. Using synthetic oligonucleotide target sequences of the maize genome, this isothermal assay has been shown to discriminate an allele with a single nucleotide polymorphism (SNP) from a wild-type. The ProxAR is designed as an end-point assay with a fluorescent read out at a fixed time point during amplification. Real-time monitoring of the reactions are performed to characterize the time-dependence of the reaction so that signal-to-noise can be optimized. The assay is amenable to correctly score allelic variations in in-bred maize lines for high-throughput detection of SNPs of interest.