GSM8965146: TC_M; Homo sapiens; RNA-Seq
RNA was harvested using a total RNA extraction kit (Tiangen Biotech). 2 μg total RNA was used for the construction of sequencing libraries. mRNAs with polyA tails were enriched by Oligo(dT) magnetic beads. The first strand of cDNA was synthesised in the 1st Strand Enzyme reverse transcriptase system using fragmented mRNA as template and random oligonucleotides as primers. The purified double-stranded cDNA was then end-repaired, A-tailed and ligated into sequencing junctions, and the 200-300bp cDNA was selected for PCR amplification and the PCR product was purified again to obtain the final library. After library construction, the libraries were quantified using a Qubit2.0 Fluorometer, diluted to 1.5 ng/ul, and the insert size of the libraries was checked using an Agilent 4200 TapeStation to ensure the quality of the libraries.