GSM6195671: MNV; Mus musculus; RNA-Seq
Small intestine (Peyer's patches removed) was cut longitudinally and rinsed in HBSS. The tissue was washed in HBSS containing HEPES, sodium pyruvate, 5 mM EDTA, and 1 mM DTT for 15 min to obtain the IEL fraction. The IEL was filtered and fractionated on a Percoll gradient (40% and 80%). The cells at the interphase of the gradient were collected and washed with complete RPMI.  Sorted IEL suspensions were loaded on a 10x Genomics Chromium instrument to generate 10,000 single-cell gel beads in emulsion (GEMs). Single-cell RNA-Seq libraries were prepared using Single Cell 3' Reagent Kits v3.1: Chromium Next GEM Single Cell 3' GEM, Library & Gel Bead Kit v3.1, PN-1000121; Chromium Next GEM Chip G Single Cell Kit, PN-1000120 and Single Index Kit T Set A PN-1000213 (10x Genomics)