muscle small RNA sequencing of Micropterus salmoides
Total RNA was isolated using TRIzol reagent (Takara, China) according to manufacturer instructions. The total RNA quantity and purity were analysis of Bioanalyzer 2100 and RNA 6000 Nano LabChip Kit (Agilent, USA) with RIN number >8.0. Approximately 1 ug of total RNA were used to prepare small RNA library according to protocol of TruSeq Small RNA Sample Prep Kits (Illumina, USA). The small RNA fraction was isolated by polyacrylamide gel electrophoresis and ligated with proprietary adaptors (Illumina, USA). Short RNAs were then reverse-transcribed into cDNA by RT-PCR. The four Small RNA libraries were constructed for M. salmoides and then sent to LC-Sciences (Hangzhou, China) for deep sequencing using an Illumina Hiseq2500 platform.