title: RNA-Seq of gallus gallus: myoblasts
design: After total RNAs were isolated from myoblasts harvested after 48 h transfection, RNA purity was assessed. Each RNA sample had an A260:A280 ratio above 1.8 and A260:A230 above 2.0. RNA integrity was evaluated using the Agilent 2200 TapeStation (Agilent Technologies, USA), and each sample had a RIN above 7.0. Briefly, rRNAs were removed from Total RNA using EpicetreRibo-Zero rRNA Removal Kit (Illumina, USA) and fragmented to approximately 200 bp. Subsequently, the purified RNAs were subjected to first strand and second strand cDNA synthesis following by adaptor ligation and enrichment with a low-cycle according to the instruction of NEBNext  Ultra RNA Library Prep Kit for Illumina (NEB, USA). The purified library products were evaluated using the Agilent 2200 TapeStation and Qubit2.0(Life Technologies, USA) and paired-end sequenced (PE150, Sequencing reads were 150 bp) at Guangzhou RiboBio CO., Ltd. (Guangzhou, China) using IIIuminaHiSeq3000.