GSM4120501: scRNAseq_Stage6C; Homo sapiens; RNA-Seq
Cells were dissociated with TrypLE, resuspended in 0.1% BSA, and filtered prior to 10X Chromium standard procedure The cells were first counted and measured for viability using the Vi-Cell XR Cell Viability Analyzer (Beckman-Coulter).  The barcoded Gel Beads were thawed from -80C and the cDNA master mix was prepared according to the manufacture's instruction for Chromium Single Cell 3' v3 library kit (10x Genomics). The live cells were mixed with the cDNA master mix.  A per sample concentration of 500,000 cells per milliliter or better was required for the standard targeted cell recovery of 2000 cells.  The stock concentration requirements would not change for higher cell recovery numbers.  The cell suspension and master mix, thawed Gel Beads and partitioning oil were added to a Chromium Single Cell B chip.  The filled chip was loaded into the Chromium Controller, where each sample were processed and the individual cells within the sample were captured into uniquely labeled GEMs (Gel Beads-In-Emulsion).  The GEMs were collected from the chip and taken to the bench for reverse transcription, GEM dissolution, and cDNA clean-up.  The resulting cDNA was a pool of uniquely barcoded molecules.  A portion of the cleaned and measured pooled cDNA continued on to library construction, where standard Illumina sequencing primers and a unique i7 Sample index were added to each cDNA pool. All cDNA pools and resulting libraries were measured using Qubit High Sensitivity assays (Thermo Fisher Scientific), Agilent Bioanalyzer High Sensitivity chips (Agilent) and Kapa DNA Quantification reagents (Kapa Biosystems).
GEO Accession
GSM4120501