RNA-Seq of Macrobrachium rosenbergii: adult hepatopancreas
mRNA sequencing libraries were prepared using NEBNext Ultra II RNA Library Prep Kit for Illumina (Cat no: NEB #E7770S) following the manufacturers instructions. Briefly, 800 ng of total RNA was used as input for poly(A) mRNA enrichment using NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB cat# E7490S) followed by fragmentation and reverse transcription to generate cDNA. Hairpin adapter was ligated to fragmented double strand cDNA and USER enzyme was used to cleave the hairpin structure. Ampure beads were used to purify adapter ligated fragments and the purified product was amplified using Illumina Multiplex Adapter primers (NEBNext Multiplex Oligos for Illumina Cat#E7500S) to generate sequencing libraries with barcodes for each sample. Library quality was checked using Agilent Bioanalyzer DNA 1000 assay kit and quantity was estimated Qubit HS DNA assay kit. The QC passed libraries were sequenced on Illumina Hiseq.