title: GSM6925423: lung metastasis infiltrating cells, scRNAseq; Mus musculus; RNA-Seq
library_construction_protocol: The lung metastatic tumors from 16 weeks MPG mice were separately collected and digested as single cells for FACS sorting by the green laser (561 nm, 100 mV). The positive fluorescent cells were discarded, and non-fluorescent cell suspension were FACS sorted out CD3 positive cells. The remaining cells after two round FACS sorting (infil) or single cells from one peritumoral lung tissue of 16 weeks MPG mice (peri) were respectively loaded onto the Chromium single cell controller (10x Genomics) to generate single-cell gel beads in the emulsion according to the manufacturer's protocol. Library was performed according to the manufacter's instructions (single cell 3' v2 protocol, 10x Genomics). Briefly, GCs were resuspended in the master mix and loaded together with partitioning oil and gel beads into the chip to generate the gel bead-in-emulsion (GEM). The poly-A RNA from the cell lysate contained in every single GEM was retrotranscripted to cDNA, which contains an Ilumina R1 primer sequence, Unique Molecular Identifier (UMI) and the 10x Barcode. The pooled barcoded cDNA was then cleaned up with Silane DynaBeads, amplified by PCR and the apropiated sized fragments were selected with SPRIselect reagent for subsequent library construction. During the library construction Ilumina R2 primer sequence, paired-end constructs with P5 and P7 sequences and a sample index were added.