title: GSM3929405: KT;H11LSL-Cas9_sgSik1#3; Mus musculus; RNA-Seq
library_construction_protocol: Tumors were dissociated using collagenase IV, dispase, and trypsin at 37C for 30 minutes. Cells were stained for CD45 (30-F11), CD31 (390), F4/80 (BM8), and Ter119 to exclude hematopoietic and endothelial cells. FACSAria™ sorters (BD Biosciences) were used for cell sorting. Total RNA was prepared from 2.50E+04 to 6.00E+04 cells using the Qiagen RNeasy micro kit. RNA quality was assessed using the RNA6000 PicoAssay kit on the Agilent Bioanalyzer 2100. Total RNA (30 ng/sample) was used for cDNA synthesis using the Ovation® RNA-Seq. Two micrograms of amplified double-stranded cDNA was sheared using a Covaris sonicator to an average length of 400 bp and subjected to library preparation using the Illumina TruSeqTM DNA sample preparation kit and sequenced (2×100 bp) on an Illumina HiSeq 4000.
identifiers.namespace: GEO
identifiers.id: GSM3929405