GSM4065488: mGst-LfngT2AVenus-SC-p01-exp1-seqrun1; Mus musculus; RNA-Seq
Gastruloids were dissociated by carefully washing them with PBS 2x, adding 50 μl Trypsin-EDTA to the well, incubating at 37 °C for 5 min 162 until they start to fall apart. Then we dissociate by pipetting up and down with a p200 pipette, inactivate Trypsin with 100 μl ESLIF, spin ~170 x g for 3 min, remove supernatant and resuspend in PBS with 10% serum. 164 Prior to FACS, DAPI (Thermo Fisher) was added to assess cell viability, and cells were sorted in the individual wells of a 384-well plate as described previously (Muraro, M. J. et al. Cell Syst 3, 385– 394.e3 (2016)) using a BD FACSJazz Cell Sorter 166 (BD Biosciences) that was equipped with BD FACS software (version 1.2.0.124). scRNA-seq on cells that were sorted was performed using the CEL-Seq2-based SORT-Seq method (Muraro, M. J. et al. Cell Syst 3, 385– 394.e3 (2016)). Bulk gastruloid A is a mix of 10 gastruloids dissociated together. Gastruloids B-J have been dissociated individually. Celseq2 runs contain the 6-nucleotide long UMI in the first nucleotides of Read 1, followed by the 8-nucleotide long cell-specific barcode. Illumina TruSeq adapaters.
GEO Accession
GSM4065488