GSM4761359: Exp120A: CD34+ cells Delta1ext-IgG hypoxia scRNA rep1; Homo sapiens; RNA-Seq
Harvested cells were resuspended at 1.0 x 105 cells/mL and encapsulated with barcoded Bead SeqB microparticles (Chemgenes) into droplets containing lysis buffer using a PDMS microfluidic device (FlowJem), aiming at capturing 3,000 - 4,000 single cells per technical replicate of each culture condition, two technical replicates were run per culture condition. Following single-cell encapsulation, droplet breakage, reverse transcription, exonuclease I treatment and PCR amplification were all performed following the standard Drop-seq protocol. The amplified cDNA and sequencing libraries were quantified using the Qubit dsDNA High Sensitivity assay (Thermo Fisher Scientific) and sized using a DNA High-sensitivity BioAnalyzer 2100 kit and instrument (Agilent Technologies). Sequencing libraries were prepared using the Nextera XT DNA sample prep kit (Illumina). The libraries were sequenced on the Illumina HiSeq 3000, with a read length of 20 bp of Read 1 using a custom primer (5' GCCTGTCCGCGGAAGCAGTGGTATCAACGCAGAGTAC 3'), 60 bp of Read 2, and 8 bp of index read 1 for sample demultiplexing. Sequencing depth was at least 50,000 raw read pairs per cell.
GEO Accession
GSM4761359