GSM4974179: DIPG-13-2; Homo sapiens; RNA-Seq
RNA was isolated using the NucleoSpin kit (Macherey Nagel) A bulk adaptation of the MARS-Seq protocol was used to generate RNA-Seq libraries for expression profiling.  30 ng of input RNA from each sample was barcoded during reverse transcription and pooled. Following Agencourct Ampure XP beads cleanup (Beckman Coulter), the pooled samples underwent second strand synthesis and were linearly amplified by T7 in vitro transcription. The resulting RNA was fragmented and converted into a sequencing-ready library by tagging the samples with Illumina sequences during ligation, RT, and PCR.