title: GSM4481179: Embryo scRNA-seq data 2; Mus musculus; RNA-Seq
library_construction_protocol: Pregnant C57BL/6 mice were sacrificed at day 9.5 post-coitum and embryos were dissected under a microscope. The yolk sack was mechanically removed from each embryo, leaving the embryo proper only, and the embryo morphology was validated to match that of a wildtype embryo of the appropriate age. To create single-cells suspension, each embryo was moved to a 1.5ml tube containing 200µl of trypsin-EDTA (0.05% trypsin, 0.02% EDTA) and incubated at 37ºC for 5 minutes. 800µl of cold MEF medium was then added to each tube to inactivate the trypsin. Single-cells were FACS-sorted directedly into a lysis solution (ddH2O, 0.1% triton-X100, RNAsin plus RNAse inhibitors) in 384-well plates. Plates were frozen and kept at -80C. lysis solution was then evaporated and reverse transcription was performed directly in the wells.  MARSseq library prepartion was performed as described in Jaitin et al. 2014 (10.1126/science.1247651). Briefly, cDNA from different cells (barcoded in the Reverse Transcripiton step) was pooled and amplified by in vitro-transcription. Amplified RNA was then fragmented, reverse transcribed, and Illumina adaptors were added by PCR (14 cycles).
attributes.tag: GEO Accession
attributes.value: GSM4481179