GSM8162812: M_medium_v2; Mus musculus; RNA-Seq
Whole pancreas ducts were isolated, seperated manually by size under a fluorescent stereoscope. Small, medium, and big ducts were further digested using 1mL of trypLE Express for 5-10min (depending on the size of the ducts) at 37ºC, vigorously shaking the digestion mix every 2-3 min. After digestion, tryPLE was neutralized by adding 10mL of blocking buffer, centrifuged, and washed twice. Sox9:eGFP positive cells were sorted from small, medoim and large ducts via FACS. Ten thousand sorted cells from each ductal fraction were pooled from 3 male mice in two independent experiments. 3' mRNA library was prepared following 10xGenomics manufacturing instructions. scRNA-seq, Sequencing was carried out as paired-end (PE77) using NovaSeq6000.