title: GSM4478171: Gastruloid 120h; Mus musculus; RNA-Seq
library_construction_protocol: Structures were serially washed through pipette transferring (cut 200ul tip) in wells filled with 200ul of 1xPBS/0.4%BSA (5 transfers). Then structures of the same time point have been pooled together and dissociated in a drop of 200 µl TrypLE Express (Gibco) for 15 minutes with pipetting every 5 minute intervals. The cell suspension was filtered using Scienceware Flowmi Cell Strainers, 40 µM. Cells were washed twice with 1 ml 1xPBS/0.4%BSA with centrifugation steps performed for 5 minutes at 1200 rpm in low DNAbind Eppendorf tubes. The cell concentration was determined using a hemocytometer and cells were subjected to single-cell RNA sequencing (10x Genomic, Chromium™ Single Cell 3' v3; one reaction per timepoint has been used) aiming for a target cell recovery of up to 10,000 sequenced cells per sequencing library (timepoint). Single-cell libraries were generated following the manual instructions of 10x Genomic (Chromium™ Single Cell 3' v3), with the exception of fewer PCR cycles in cDNA amplification and sample index PCR than recommended in order to increase complexity of libraries. Libraries were sequenced with a minimum of 230 Mio paired end reads and parameters as described in the manual.
attributes.tag: GEO Accession
attributes.value: GSM4478171