GSM3897992: Sorted_cells_pool_4; Mus musculus; RNA-Seq
To dissociate the enteroids, the culture medium was removed and the Matrigel was broken. Then, the enteroids were digested to single cells in 500µL of TrypLE™ Express Enzyme (1X) (Gibco™) at 37°C for 20min. The dissociation was improved by passaging into a syringe mounted with a 18G needle. The digestion was stopped by addition of Advanced DMEM/F-12, fetal calf serum 10%, N-acetylcysteine 0,5mM, B27 1X. The cells were pelleted, washed 2 times and resuspended in the same medium. Finally, they are filtered on a 50 µm cell strainer. eYFP+ cells, from dissociated Neurog3eYFP/+ enteroids, were sorted directly into 96 well plates « Precise WTA Single Cell Encoding Plate » (BD™ Precise WTA Single Cell Kit, BD Genomics) (1 cell/well) using a FACS Aria Fusion (BD). DAPI was used to sort only living cells. Cells were sorted directly into four 96 well plates « Precise WTA Single Cell Encoding Plate » (BDTM Precise WTA Single Cell Kit, BD Genomics) (1 cell/well) using a FACSAria II (BD). The library was prepared by the GenomEast Platform (IGBMC) following the protocol « Library Preparation with the BDTM Precise WTA Single Cell Kit User's Guide » (910000014 Rev. 02 11/2016, BD Genomics).
GEO
GSM3897992