cDNA of Thunnus thynnus: muscle tissue (TunaWMED04)
Starting from total RNA, mRNA was obtained with the mRNA Isolation Kit (Roche) from 10 individuals (3 from Western Mediterranean, 3 from Eastern Mediterranean and 4 from Gulf of Mexico) and used as template for cDNA libraries synthesis according to Clontech's SMARTer cDNA kit. A addition a modified oligo-dT primer (5-AAGCAGTGGTATCAACGCAGAGTTTTCTTTTTTCTTTTTTV3) was used for first strand synthesis as described in (Beldade et al. 2006; Meyer and Kircher 2010). Libraries were purified with the QiaQuick PCR purification kit (Qiagen) and quantified, then processed according to the Roche protocol for cDNA Rapid Library preparation. Sequencing was performed on a full plate run of the Roche GS-FLX DNA Sequencer with Titanium chemistry