GSM8776407: MCF7, paclitaxel-resistant, 2; Homo sapiens; RNA-Seq
Cultured cells were lysed with 1 ml TRIzol (Ambion), and after adding 200 µl chloroform, the samples were vortexed and centrifuged at 12000 rpm for 15 min at 4 °C. The supernatant was mixed with 30 µl NaOAc, 0.5 µl glycoblue, and 500 µl IPA, then incubated overnight at -20 °C. The pellet was collected by centrifugation at 15000 rpm for 1 h at 4 °C, washed twice with 75% EtOH, dried, and dissolved in 42 µl TDW. For DNA removal, 2 µl DNase I (Takara), 1 µl RNase inhibitor, and 5 µl DNase buffer were added and incubated at 37 °C for 30 min. The sample was purified with 150 µl TDW and 200 µl acid phenol, followed by ethanol precipitation overnight. The pellet was washed with 75% EtOH, dried, and dissolved in TDW. RNA libraries for RNA-seq was prepared using Illumina's TruSeq stranded mRNA LT Sample Prep Kit following the manufacturer's protocols.