title: GSM4773461: mCorpus WT rep1; Mus musculus; RNA-Seq
library_construction_protocol: Freshly isolated gastric epithelium was isolated according to Bartfeld et al. 2015. In short, gastric human biopsies or mouse tissue regions were cut into pieces and incubated rotating in chelating solution at 4 °C for 30 min. Thereafter, gastric glands were squeezed out of the tissue pieces by applying pressure on a glass slide, which was placed on top, and collected for single cell digestion for 5 min in TrypLE solution. The resulting single cell suspension was then used for FACS. Living single cells (DAPI negative) were sorted in 384 well plates. The RNA of single cells was processed according the CEL-seq2 protocol (Muraro et al., 2016). In short, total RNA of single cells was reverse transcribed into cDNA in the 384 well plate, thereafter pooled and in vitro transcribed by T7 polymerase overnight. After fragmentation of the amplified RNA, it was once more reverse transcribed before library preparation for next generation sequencing was performed. The quality of the fragmented RNA and cDNA library was check by PICO RNA or DNA hypersensitivity chip (Agilent), respectively. The samples were then paired-end sequenced on NextSeq500 (2x 75bp) by the Utrecht Sequencing Facilities (www.useq.nl).