Wu10
Cells collected and enriched for viable cells to greater than 90% using the dead cell removing kit (Miltenyi Biotec, Cat#130-090-101), submitted for cell capturing and single cell RNA sequencing. Barcoded Gel Beads were thawed from - 80C to room temperature and the cDNA master mix was prepared according to the manufactures instruction for Chromium Single Cell 3 v2 library kit (10x Genomics). The target capture number of cells was 4000, so a volume containing more than 5000 live cells was combined with cDNA master mix. The cell suspension and ChromiumSingle Cell 3 v2 library master mix (10x Genomics), 492 gel beads, and partitioning oil were added to a Chromium Single Cell A chip which was then loaded into the Chromium Controller for capturing and cDNA synthesis. The resulting cDNA libraries were pooled and sequenced as 100x2 paired end reads on an Illumina HiSeq 4000 using HiSeq 3000/4000 sequencing kit.