GSM6689467: MCF-7_Nutlin-3a_1; Homo sapiens; RNA-Seq
Total RNA was extracted using the RNeasy Plus Mini Kit (Qiagen) following the manufacturer's protocol. Libraries were prepared from 800 ng of input material (total RNA) using NEBNext Ultra II Directional RNA Library Preparation Kit in combination with NEBNext Poly(A) mRNA Magnetic Isolation Module and NEBNext® Multiplex Oligos for Illumina®(Index Primers Set 1/2/3/4) following the manufacturer's instructions (New England Biolabs) including size-selection at around 500 bp. Quantification and quality checked of libraries was done using a 2100 Bioanalyzer instrument and DNA 7500 kit (Agilent Technologies). Libraries were pooled and sequenced on a NextSeq 500 v2 300 cycles runs. System run in 151 cycle/paired-end mode.