Coding sequences were screened for polymorphisms in 163 UK individuals with schizophrenia and 135 UK blood donor controls. Genomic sequences corresponding to all exons were extracted from the UCSC Genome Browser (March 2006), based upon Refseq transcript NM_194250. Amplicons with a maximum length of 500 bp (n=13) were designed to span coding exons plus a minimum of 72 bases of flanking sequence. Where multiple amplicons were required to cover an exon, these overlapped by a minimum of 81bp. High Resolution Melting Analysis (HRMA) was performed for 11 amplicons using a LightScanner™ (Idaho technologies) according to the manufacturer’s instructions. Potential DNA variants identified by this approach were characterised by sequencing the relevant PCR product using BigDye chemistry on a 3100 capillary sequencer (Applied Biosystems). Two amplicons failed to optimise in the presence of the HRMA detection reagent and were screened for variants by sequencing. Identified SNPs were genotyped in the CEU HapMap trios to assess their LD relationships, and to detect Mendelian inconsistencies (of which there were none).
