title: GSM8141547: 3CellLines scComplete-seq GEX 1XSPRI; Homo sapiens; OTHER
library_name: GSM8141547
library_construction_protocol: Single cells of 3 cel lines and PBMCs were encapsulated in Gel-Bead-Emulsions (GEMs) using the 10X Controller with a Chromium Single Cell 3' GEM Library and Gel Bead Kit (Singleron, Singapore) The cell lines after the aspiration of the supernatant, the cells were washed in washing buffer.  Phosphate buffered saline (PBS) with 0.02% FBS was used as the staining and washing buffer. K562, MCF7 and HEK293T cells were then blocked with Fc-blocking agent (BioLegend, 422301) at 4 °C for 30 min and labeled with sample identifier hashtags (0.5 µg each Ab-oligo per million cells, TotalSeq™-A0251/2/3 anti-human Hashtag 1/2/3). Cells were washed with wash buffer 3 times and pooled together (equal number of cells for each, loading 4,800 cells in total). PBMCs from different conditions were then blocked with Fc-blocking agent and labeled with sample identifier hashtags given above (0.5 µg each Ab-oligo per million cells) at 4 °C for 30 min. Cells were washed with wash buffer 2 times and pooled together (equal number of cells per condition, and loading 16,000 cells in total). An Ab-oligo cocktail (0.125 µg each Ab-oligo per million cells, TotalSeq™- A anti-human CD3, CD4, CD8, CD16, CD27, CD28, CD45, CD45RA, CD56, CD69, CD95, CD197) was prepared in 45 µl of staining buffer and 5 µl  of Fc-blocking reagents immediately before use. The pooled barcoded cell samples were stained with surface marker Ab-oligo cocktail at 4 °C for 40 min. Cells were washed with wash buffer 3 times and resuspended in PBS.