GSM4119369: CDM day 6; Homo sapiens; RNA-Seq
Cells were released enzymatically from the culture flasks by incubation with TrypLE Express (Gibco, cat #12605-036) for 10 minutes, spun down at 200g for 10 minutes and resuspended at a cell density of 1 million cells/ml in 0.1% BSA-PBS. Libraries were prepared using the Chromium Single Cell 3' Reagent Kit (v2 Chemistry, 10x Genomics) according to the manufacturer's instructions. Briefly, Gel bead-in-Emulsions (GEMs) were generated using the Chromium Single Cell Controller. Following reverse transcription, the single-cell droplets were broken. Single-strand cDNA was purified with Cleanup Mix containing Silane DynaBeads (Thermo Fisher Scientific) and subsequently amplified. Next, the SPRIselect Reagent Kit (Beckman Coulter) was used for fragmentation, end-repair, A-tailing and index adaptor ligation. Final libraries were obtained after amplification and subsequent clean-up of the ligated strands with SPRIselect beads. Sequencing of the libraries was performed on the HiSeq4000 System (Illumina) using the following parameters: 26 bp read 1 – 8 bp index 1 – 98 bp read 2
GEO Accession
GSM4119369