title: Single-Cell RNA-Seq GM12878
design: Lymphoblastoid cell lines obtained from the Coriell Cell Repositories were established by Epstein-Barr Virus (EBV) transformation of peripheral blood mononuclear cells using phytohemagglutinin as a mitogen.  Cells were cultured in the recommended Roswell Park Memorial Institute (RPMI) Medium 1640 supplemented with 2mM L-glutamine and 20% of fetal bovine serum on T25 tissue culture flask with 20 mL medium upright position at 37C under 5% carbon dioxide. Lymphoblast cultures where split every four days to yield a cell density of 400,000 - 600,000 viable cells/mL. Suspended cells were pelleted by centrifugation in PBS (400g, 10min, 4C), the supernatant was discarded, and the pellet was resuspended in 1mL ice-cold PBS. Cell numbers and viability were assessed by Trypan blue staining and counting in a Neubauer improved counting chamber. Libraries were prepared using the Chromium controller (10X Genomics, CA) in conjunction with the single-cell 3 v2 kit. Briefly, the cell suspensions were diluted in nuclease-free water according to manufacturer instructions to achieve a targeted cell count of 5000. cDNA synthesis, barcoding and library preparation were then carried out according to the manufacturer's instructions. The libraries were sequenced on an Illumina Novaseq 6000 (Illumina, San Diego) with a read length of 26 bp for read 1 (cell barcode and unique molecule identifier (UMI)), 8bp i7 index read (sample barcode) and 98 bp for read 2 (actual RNA read).