Rna-seq of homo sapiens: 3_CTRL
After the total RNA QC procedures, the mRNA isolated from human mesenchymal stem cells was enriched for polyA RNA (mRNA) using oligo(dT) beads. After enrichment, the mRNA was fragmented randomly by adding fragmentation buffer, then the cDNA was synthesized by using mRNA template, random hexamers primers. The custom second-strand synthesis buffer (Illumina), dNTPs, RNase H and DNA polymerase I were added to initiate the second-strand synthesis. After the series of terminal repair, adaptors ligation, the double-stranded cDNA library was completed through size selection and PCR enrichment. The qualified libraries were pooled and fed into Illumina sequencer, according to their effective concentration and expected data volume. The polyA transcriptome libraries were sequenced on HiSeq4000 (Illumina, San Diego, CA, USA) sequencer with the following parameters: PE 150 (paired ends) and 40M clean reads which gave a minimum of 12 Gb of raw data per each sample.