GSM8634247: MDA-MB-231 tramadol 0 mg/ml; Homo sapiens; RNA-Seq
RNA was extracted using the RNeasy kit (Qiagen, 74104) following the manufacturer's instructions. For RNA-seq, 1 μg of total RNA was used for library preparation. Poly(A) mRNA isolation was performed using Oligo(dT) beads. mRNA fragmentation was performed using divalent cations and high temperature. Priming was performed using random primers. First-strand cDNA and second-strand cDNA were synthesized. The purified double-stranded cDNA was then treated to repair both ends, and dA-tailing was added in one reaction, followed by T-A ligation to add adaptors to both ends. Size selection of adaptor-ligated DNA was then performed using DNA Clean Beads. Each sample was then amplified via PCR using the P5 and P7 primers, and the PCR products were validated. Libraries with different indices were then multiplexed and loaded on an Illumina HiSeq/Illumina Novaseq/ MGI2000 instrument for sequencing using a 2x150 paired-end configuration according to the manufacturer's instructions.