GSM7719458: Human Lung DTC rep 1 [Seagen]; Homo sapiens; RNA-Seq
Frozen samples of primary human lung tumors underwent dead cell removal and T cell enrichment using the Sony MA900 Cell Sorter with a DAPI live/dead cell discriminator and staining for CD45 and CD3. Following dead cell removal and T cell enrichment (CD45+ 972 CD3+), cell count and viability were assessed using acridine orange/propidium iodide on a Nexcelom Cellometer and cells were washed twice in PBS + 0.04% bovine serum albumin. Following successful isolation, 15,000 cells per sample were loaded into the 10X Chromium chip using the Chromium Single Cell (v3.1 chemistry) with the intention of targeting 10,000 cells/sample for sequencing. 3′ GEX scRNA-Seq libraries were prepared according to manufacturer recommendations. Libraries were quality controlled on the Agilent TapeStation. Libraries were pooled and sequenced on the NovaSeq 6000 S2 100 flow cell targeting an average depth of 25,000 reads per cell (×10,000 cells/sample) or 250 million reads/sample.