Primers used for analysis of four SNPs in one gene.

The primers in (S1 Table) were generated by the design software, guided by Agena MassARRAY® system. 4 SNPs were designed into 3 pools. The primary polymerase chain reaction (PCR) that is locus-specific PCR was amplified with pools of 1st PCRP and 2nd PCRP for specific SNP loci. This secondary PCR uses mass-modified dideoxy nucleotide terminator of an oligonucleotide primer (UEP_SEQ). Primer anneals immediately upstream of the polymorphic site of interest. The SNPs were added in a multiplexed single base pair extension (SBE) with dideoxy nucleotides that are mass modified depending on the allele and design of the assay. The extended primers were then detected by MALDI-TOF MS in the analyzer.
(XLSX)