GSM3408328: cervical intraepithelial neoplasia 4; Homo sapiens; RNA-Seq
PAXgene blood RNA tubes were used to collect 2.5ml blood. Total RNA was extracted using the PAXgene Blood RNA Kit. NEBNext® Ultra RNA Library Prep Kit was used with 2 ug of total RNA for the construction of sequencing libraries. Sequencing libraries were generated using NEBNext® Ultra RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer's recommendations. Briefly, mRNA was captured from 2μg total RNA using poly-T oligo-attached magnetic beads. Fragmentation was carried out using divalent cations under elevated temperature to make mRNA approximate 200 bp inserts. First and second strand cDNA were synthesized, then purified using AMPure XP beads. After end repair, an Illumina adapter was ligated to the sticky 5'end, then different i5 and i7 primers were added to each sample library, following 12 cycles of linear PCR amplification.