GSM8878744: Control oligo; Mus musculus; RNA-Seq
The ascite cells were resuspended in digestion buffer (HBSS withMg2+/Ca2+ supplemented with 2 mgml collagenase type l and 40 U/ml DNase l)and incubated at 37 'C for 30 min. Cells were washed and re-suspended in PBS and stained with a Viability Dye.ascite cells were washed in HBSS-/and resuspended in HBSS-/supplemented with 0.04% w/ BSA. Library was performed according to the manufacter's instructions (single cell 3' v2 protocol, 10x Genomics). Briefly, GCs were resuspended in the master mix andoaded together with partitioning oil and gel beads into the chip to generate thegel bead-in-emulsion (GEM). The poly-A RNA from the cell lysate contained inevery single GEM was retrotranscripted to cDNA, which contains an llumina Riprimer sequence, Unique Molecular ldentifer (UMl) and the 10x Barcode. Thepooled barcoded cDNA was then cleaned up with Silane DynaBeads, amplifedby PCR and the apropiated sized fragments were selected with SPRlselectreagent for subsequent library construction. During the library constructionllumina R2 primer sequence, paired-end constructs with P5 and P7 sequencesand a sample index were added.