GSM9081868: 5442I; Mus musculus; RNA-Seq
The jejunum and ileum of a P6 male and female mouse of was isolated and rinsed with HBSS. The tissue was segmented into ~2 cm fragments and opened longitudinally to expose the epithelium. The tissue was incubated in 30 mM EDTA in HBSS without calcium and magnesium at 37˚C for 20 min. The tissue was vigorously shaken to release epithelium. The epithelium was collected, incubated on ice, and allowed to settle via gravity. The epithelium was washed twice in HBSS, then transferred to 1 mg/ml dispase II in HBSS and triturated for 10 passes. Cell suspension was incubated on macs dissociator for 12 mins at 200 rom at 37C.  Cells were strained and washed with HBSS +10% FBS and centrifuged at 300 G for 5 mins. Cells were centrifuged and washed in cell suspension buffer (PIPSeq kit) and cells were resuspended at a concentration of ~4000 cells/uL. Single cell libraries were prepared using PIP-Seq V T20 kit