title: GSM1931300: J1015_treated; Homo sapiens; RNA-Seq
library_construction_protocol: Five slides (5 µm each) from frozen xenografts were used for total RNA extraction using TRIzol reagent (Invitrogen). RNA was purified using a miRNeasy kit (Qiagen) according to the manufacturer's instructions. RNA-seq transcriptome libraries were prepared following the TruSeq RNA Sample Preparation v2 (Illumina, Part# 15026495 Rev. A), using 2 μg of total RNA. PolyA+ RNA was isolated using Sera-Mag oligo(dT) beads (Thermo Scientific) and fragmented with the Ambion Fragmentation Reagents kit. cDNA synthesis, end repair, A-base addition and ligation of the Illumina-indexed adaptors were performed according to Illumina's protocol. Libraries were then PCR amplified using Phusion DNA polymerase (NEB) for 14 PCR cycles before quantification and multiplexing. 10 pmol of multiplexed (n=4) paired-end libraries were sequenced with the Illumina HiSeq1000 (2 × 50-nt read length). Reads that passed the chastity filter of Illumina BaseCall software were used for subsequent analysis. RNA libraries were prepared for sequencing using standard Illumina protocols
attributes.tag: GEO Accession
attributes.value: GSM1931300