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We attempted to overcome these limitations by focusing on a single pathway, using an information-based gene-selection algorithm (based on location in autism linkage regions and expression patterns), selecting tag-single-nucleotide polymorphisms (SNPs) to achieve very high coverage of genes and flanking regions, and by evaluating high order genegene interactions using a newly developed method adapted for family data

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RT-PCR was performed using LightCycler 480 II Real-time PCR Instrument (Roche, Basel, Switzerland) with QuantiFast SYBR Green qPCR Master Mix (Qiagen, Dsseldorf, Germany)