GSM6339377: NDV-infetion HD11 cells rep3; Gallus gallus; RNA-Seq
Total RNA from HD11 cells was extracted at 24 hpi using TRIzol reagent (Invitrogen Corporation, CA, USA) according to the manufacturer's instruction. RNA concentration and quality of each sample were determined to measure the OD260/OD230 value and OD260/OD280 value by NanoDrop ND-1000 (Thermo Fisher Scientific, Waltham, MA, USA). RNA integrity and gnomic DNA contamination were tested by denatured agarose gel electrophoresis. MeRIP‑Seq service was performed by Cloudseq Biotech Inc (Shanghai, China). Briefly, RNA from each sample was separately chemically fragmented into fragments with a length of approximately 100 nucleotides using fragmentation buffer (Illumina Inc.). Immunoprecipitation of fragmented RNA was performed with the GenSeq™ m6A-MeRIP Kit (GenSeq Inc., China) according to the the manufacturer's instructions. Both the input samples without immunoprecipitation and the m6A immunoprecipitation samples were used for RNA-seq library construction with NEBNext® Ultra II Directional RNA Library Prep Kit (New England Biolabs, Inc., USA) following the manufacturer's instruction. The library quality was evaluated with BioAnalyzer 2100 system (Agilent Technologies, Inc., USA). Library sequencing was subjected to 150 bp paired-end sequencing on an Illumina NovaSeq 6000 instrument.