GSM8777872: LNCaP_C4-2B, steady state grown in RPMI1640 supplemented with 10%FBS and 1%Pen/Strep antibiotic.; Homo sapiens; RNA-Seq
To collect single cell suspension, cells were trypsinized and washed once in PBS and re-suspended in cell culture media followed by haemocrytometer cell count to ensure cell viability above 70%. The cell suspension were loaded into Chromium Next GEM Chip G (10x Genomics , PN#1000120) with 3′ v.3.1 beads (10x Genomics , PN#2000164) and partitioning oil.  Individual cells were barcoded using a 10× Chromium Controller (10x Genomics). RNA from the barcoded cells were subsequently reverse-transcribed and amplified with a Chromium Single Cell 3′ v3.1 Reagent Kit (10x Genomics, PN#1000123).  Libraries were subsequently constructed from 25% of amplified cDNA traces from pre-vious step with reagents from a Chromium Single Cell 3′ v3.1 Library Kit (10x Genomics, PN#1000190). The cDNA and library traces were quantitated and identified by Tapestation.  Sequencing were performed with an Illumina NextSeq 2000 P3.