Full length transcritome of noble scallop
mRNA was purified from 15 g of mixed total RNA using poly (T) oligo-attached magnetic beads. Fragmentation was conducted using divalent cations under elevated temperatures in the NEBNext First Strand Synthesis Reaction Bufer (5). The SMART PCR cDNA Synthesis Kit (Clontech, CA, USA) was used for synthesizing full-length cDNA. Remaining overhangs were converted into blunt ends via exonuclease/polymerase activities. After adenylation of the 3 ends of the DNA fragments, NEBNext Adaptor with a hairpin loop structure was ligated to prepare for hybridization. BluePippin (Sage Science, Beverly, MA, USA) was used for size selection of the full-length cDNA and for building libraries of differently sized cDNA. The generated cDNA was then re-amplified using PCR, and the fragment size distribution was quantified using the Qubit fuorometer (Life Technologies, Carlsbad, CA, USA). The quality of the libraries was assessed using the Agilent Bioanalyzer 2100 system. SMRT sequencing was performed using the Pacific Biosciences real-time sequencer using C2 sequencing reagents.