title: GSM7025157: HSJD-DIPG-007-DMSO-1; Homo sapiens; RNA-Seq
library_construction_protocol: RNA extraction was carried out using the RNeasy Mini kit (Qiagen), including on-column DNase I digestion (Invitrogen). Integrity of genomic DNA was confirmed by gel electrophoresis. RNA was assessed on an Agilent 2100 Bioanalyzer using the Agilent RNA 6000 Nano kit. Strand specific mRNA libraries were generated using the NEBNext Ultra II Directional RNA library prep Kit for Illumina (New England BioLabs #E7760), mRNA was isolated using Poly(A) mRNA magnetic isolation module (New England BioLabs #E7490). Preparation of libraries followed the manufacturer's protocol (Version 1.0 4/17).  Input was 1000ng and samples were fragmented for 15 min for RNA insert size of ~200 bp.  The following PCR cycling conditions were used: 98°C 30s / 8 cycles: 98°C 10s, 65°C 75s / 65°C 5 min. Stranded mRNA libraries were sequenced on an Illumina HiSeq4000 instrument,  using 47bp paired-end dual indexed reads and 1% PhiX control.