Pinctada maxima pooled cDNA 454-EST database for SNP discovery
To enable genome-wide SNP discovery, high quality total RNA was extracted from mantle tissue of 62 P. maxima individuals using TRIZOL® Reagent (Life Technologies). Individual samples were pooled together in equimolar amounts. Full-length double stranded cDNA was produced using the Mint cDNA synthesis kit (Evrogen) and normalised using Trimmer cDNA normalisation kit (Evrogen) using manufacturer’s recommendations. cDNA was normalised to promote the sequencing of rare and unique gene transcripts. Normalised cDNA was pyrosequenced using 454 FLX technologies (Roche) according to manufacturer’s protocols.