mRNA-seq of Pelteobagrus vachelli muscle (M4b)
Total RNA was extracted using Trizol reagent (Invitrogen, CA, USA) following the manufacturers protocol. The total RNA quantity and purity were analyzed with Bioanalyzer 2100 and RNA 6000 Nano LabChip Kit (Agilent, CA, USA) with RIN number >7.0. For six cDNA library constructions (M0a, M0b, M0c, M4a, M4b, M4c), Poly(A) RNA is purified from total RNA(5ug) using poly-T oligo-attached magnetic beads using two rounds of purification. Following purification, the mRNA is fragmented into small pieces using divalent cations under elevated temperature. Then the cleaved RNA fragments werereverse-transcribed to create the final cDNA library in accordance with the protocol for the mRNA-seq sample preparation kit (Illumina, San Diego, USA), the average insert size for the paired-endlibraries was 300 bp (50 bp). And then we performed the paired-end sequencing on an Illumina Hiseq 4000 at the (LC Sceiences,USA) following the vendors recommended protocol.
SUB7765255
M4b