title: RIP_seq-R1_WT_r1
design: RNA immunoprecipitation was performed with reversible cross-linking of the cells, as previously described (Niranjanakumari et al., 2002). The RNA was extracted by using Trizol reagent (Invitrogen) .RNA quality was determined. High-quality RNA sample is used to construct sequencing library. RNA-seq transcriptome libraries were prepared following TruSeqTM RNA sample preparation Kit from Illumina (San Diego, CA). cDNA synthesis, end repair, A-base addition and ligation of the Illumina-indexed adaptors were performed according to Illumina protocol. Libraries were then size selected for cDNA target fragments on 2% Low Range Ultra Agarose (Sigma-Aldrich) followed by PCR amplified using Phusion DNA polymerase (NEB) for 15 PCR cycles. After quantification, paired-end libraries were sequenced by Illumina NovaSeq 6000 sequencing