title: GSM4576006: HG3, Low survivorship - timepoint 2; Homo sapiens; RNA-Seq
library_construction_protocol: Forty million cells from the bulk TP0 and TP1 populations were co-stained using human CXCR4-APC (Thermo Fisher #FAB173A) and CD18-PE (BioLegend #302108) conjugated antibodies. CXCR4 and CD18 single-positive cells (representative of the high and low survivorship sub-populations) were isolated using a BD FACSAria II. Three biological replicates were collected in this manner over the course of two weeks. RNA was isolated from purified populations using a RNeasy Micro Kit (Qiagen #74004) and RNA quality was verified using an Agilent Bioanalyzer Pico Eukaryote chip. 400 ng of total RNA was used as input for library preparation using the KAPA stranded mRNA HyperPrep Kit (KAPA Biosystems #KK8580) per the manufacturer's instructions. Library yields and sizes were confirmed using an Agilent High Sensitivity DNA kit (Agilent Technologies #5067-4626) on an Agilent 2100 bioanalyzer. Libraries were pooled in equimolar ratios and sequenced using a 75-cycle high output kit on an Illumina NextSeq 500 system. 