LIVER TRANSCRIPTOME AND SNP DISCOVERY IN THE FISH Piaractus mesopotamicus
A non-normalized cDNA library was prepared using cDNA Synthesis System Kit with random primer GS Rapid Library Prep Kit and GS Rapid Library MID Adaptors Kit (Roche). We used the High Sensitivity DNA LabChip Kit (Agilent Technologies) with 2100 Bioanalyzer for quality analysis of the cDNA library.The concentration of sample molecules/uL was obtained by QuantiFluorTM –ST fluorimeter (Promega). Titration of emPCR (emulsion PCR) was performed with the GS FLX Titanium SV em PCR Kit (Lib-L) (Roche), according to the emPCR Amplification Method Manual – Libl SV, GS FLX+ Series, to identify the optimal number of DNA molecules per bead (cpb = copies per bead). After emPCR titration, the emPCR was performed with GS FLX Titanium LV emPCR Kit (Lib-L) (Roche), according to the emPCR Amplification Method Manual – LibL LV, GS FLX+ Series. The transcriptome sequencing was done with the Roche/454 technology (GS FLX Titanium Sequencing Kit XL +).