rawdata.rar

Total RNA (1 μg) of GC cells were used to prepare small RNA libraries by NEBNext Multiplex Small RNA Library Prep Set for Illumina (NEB, USA) according to the manufacturer’s instructions. The libraries were sequenced by HiSeq 2500 (Illumina, USA) with single-end 50 bp at RiboBio Co. Ltd (RiboBio, China). The raw reads were processed by filtering out the ones containing adapter, poly ’N’, of low quality, and which were smaller than 17nt reads by FastQC to get clean reads. Mapping reads were obtained by mapping clean reads to the reference genome of BWA software. miRDeep2 was used to identify known mature miRNA based on miRBase21 (www.miRBase.org) and predict novel miRNA. Databases of Rfam12.1 (www.rfam.xfam.org) and piRNABank (www.pirnabank.ibab.ac.in) were used to identify ribosomal RNA (rRNA), transfer RNA (tRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), and piwi-interacting RNA (piRNA) by BLAST. The miRNA expression were calculated in RPM (reads per million) values (RPM = (number of reads mapping to miRNA/ number of reads in clean data) × 10⁶). The expression levels were normalized by RPM [(number of reads mapping to miRNA/number of reads in clean data) × 10⁶].