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Primers were designed with the Primer3 software http://www-genome.wi.mit.edu/cgi-bin/primer/primer3_www.cgiand were synthesized by Sigma-Genosys.
A user interface is available at http://gene.cpmc.columbia.edu/cgi-bin/gene.cgi.
Once a sequence containing the desired polymorphism was selected (usually 400-700 bp in length), it was run through the Repeat Masker program http://ftp.genome.washington.edu/cgi-bin/RepeatMaskerin order to avoid selecting PCR primers within Alu, L1, or other repeats.
Intronic oligonucleotide primer pairs flanking the coding fragments were designed for both the genes using program primer 3 http://www.genome.wi.mit.edu/cgi-bin/primer/primer3.cgiusing the genomic sequences of SLC15A1 and GPC5 gene respectively.
The gene was located in the approximate interval both cytologically and on the Santa Cruz map http://genome.ucsc.edu/cgi-bin/hgGateway?db=hg10, but was located on chromosome 16p on the NCBI map (builds 25 to 28).