MARS-seq Version 2

ABSTRACT: scRNA-seq procedure and analysis. ScRNA-seq was performed using the Massively Parallel Single-Cell RNA-sequencing technology (MARS-seq) as described by Jaitin et al., 2014 (PMID: 24531970) EpiSCs were seeded at ~6 x 103 cells/cm2 in 6-well cell-culture plates pre-coated with 15 µg/ml human Fibronectin, and differentiated to APSD. At desired time-points, cells were dissociated using Accutase (Thermo Fisher Scientific, 00-4555-56) for 3 min at 37°C and counted. 500,000 cells per condition were washed with ice-cold FACS buffer (10% (v/v) FBS in PBS) and resuspended into 1 ml ice-cold FACS buffer containing 1 µg/ml DAPI. For in vivo experiments, E6.5/E7.0/E7.75 Tg(Eomes::GFP) BAC transgenic embryos were individually dissociated into 300 μl warmed Trypsin 0.05% (w/v) EDTA for 10 min at 37°C.

INSTRUMENT(S): NextSeq 500, Bravo automated liquid handling robot station (Agilent)

ORGANISM(S): Mus musculus

SUBMITTER:  

PROVIDER: E-MTAB-13660 | biostudies-arrayexpress |

SECONDARY ACCESSION(S): ERP156294

REPOSITORIES: biostudies-arrayexpress