title: GSM7882929: Dox_Exos, replicate 1, scRNAseq; Homo sapiens; RNA-Seq
library_construction_protocol: The adherent LO2 cells were digested with 0.25% trypsin for 4 minutes and digestion was terminated by adding high-glucose DMEM (GIBCO) supplemented with 10% fetal bovine serum (FBS) (GIBCO). The dissociated cells were centrifuged at 1,000 rpm and 4 °C for 5 min, then resuspended in 15 mL red blood cell lysis buffer (BD Biosciences) at 37 °C for 5 min. Subsequently, these cells underwent another round of centrifugation at 1,000 rpm and 4 °C for 5 min, followed by two washes with cold PBS and resuspended in cold PBS containing 10% PBS. The cell suspension was filtered through a 40 μm strainer (BD Falcon), subjected to two PBS washes, centrifuged at 1,000 rpm and 4 °C for 5 min, and resuspended in cold PBS supplemented with 10% FBS. The library was performed according to the manufacturer's instructions (single cell 3' v2 protocol, 10ÍGenomics). 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 emulsion (GEM). The poly-A RNA from the cell lysate contained in every single GEM was retro-transcripted to cDNA, which contains an Ilumina R1 primer sequence. Unique Molecular Identifier (UMI) and the 10ÍBarcode. The pooled barcode cDNA was then cleaned up with Silane DynaBead, 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 sequence P5 and P7 sequences and a sample index were added.