Equine ASIP Gene Sequencing

Reference genomic sequences were extracted from Ensembl (Equine ASIP gene, ENSECAG00000004241). PCR and sequencing primers were designed using Primer3 [13 (link)]. The three exons were amplified using three sets of primers [See Additional file 1: Table S1]. PCR amplicons were sequenced using Sanger dideoxy sequencing in both forward and reverse directions by GATC Biotech (GATC Biotech AG, Konstanz, Germany). Electropherograms were manually inspected with Chromas Lite (Technelysium Pty Ltd, South Brisbane, Australia). Multiple alignments were performed using Multalin ([14 (link)]; http://multalin.toulouse.inra.fr).