title: GSM2148796: Single-cell RNA-seq in EB+36hrDox(iNgn2.Isl1.Lhx3.V5) cell 2; Mus musculus; RNA-Seq
library_construction_protocol: Cells were collected at different time points after NIL induction. Differentiating embryoid bodies were washed with phosphate-buffered saline and then dissociated by mild trypsinization followed by mechanical dissociation into single cell suspension. Viable cells were labeled by incubating with 1 µM Fluorescein diacetate (Sigma) at 37°C for 10 minutes. Single cells were FACS-sorted into 96-well plates containing 10 µl of 1% beta-mercaptoethanol in TCL buffer (Qiagen). Once sorting was completed, plates were sealed, centrifuged 1 min at 800 g at RT and immediately frozen on dry ice. Plates were kept at -80°C until lysate cleanup and reverse transcription of mRNA. Single cell libraries were prepared in two overall batches using a custom version of the SMART-Seq2 protocol (Picelli et al., 2013), as previously described (Satija et al., 2015), without the use of Random Molecular Tags. Cells were sequenced with 50bp single end reads on a HiSeq 2500 at an average sequencing depth of 325,000 reads per cell.
attributes.tag: GEO Accession
attributes.value: GSM2148796