GSM9049493: UEIS LSL; Mus musculus; RNA-Seq
The MC38 tumour tissue grown in UEISLSL and UEISKI mice were dissected and digested into single-cell suspension with 0.1 mg/ml DNAase I, and 0.6 mg/ml Collagenase II at 37 °C for 30 min. The cells were filtered through a sterile 70 µm cell strainer to remove red blood cells. Library preparation was performed following the manufacturer's instructions (Single Cell 3' v3 protocol, 10x Genomics). Briefly, GCs were resuspended in the master mix and loaded with partitioning oil and gel beads into the microfluidic chip to generate gel bead-in-emulsions (GEMs). Within each GEM, poly-A RNA from lysed cells was reverse-transcribed into cDNA containing an Illumina R1 primer, a Unique Molecular Identifier (UMI), and a 10x barcode. The barcoded cDNA was then purified using Silane DynaBeads, amplified by PCR, and size-selected with SPRIselect reagent. During library construction, Illumina R2 primer sequences, paired-end adapters (P5 and P7), and sample indices were added.