Sample 9
Mus musculus
Protocols: Outbred CD1 females were timed mated to generate mouse embryos of the specified stages. The morning that the vaginal plug was detected was defined as embryonic stage (e)0.5. For neural tube dissection, cervical and thoracic sections of single mouse embryos were dissected in Hanks Balanced Solution without calcium and magnesium (HBSS, Life Technologies 14185045) supplemented with 5% fetal bovine serum (FBS). The samples were then incubated on FACSmax cell dissociation solution (Amsbio T200100) with 10X Papain (30U/mg Sigma10108014001) for 11min at 37°C to dissociate the cells. To generate a single cell suspension, samples were transferred to HBSS, with 5%FBS, rock inhibitor (Y-27632 10uM, Stem Cell Technologies), and 1X non-essential amino acids (11140035), disaggregated through pipetting, and filtered once in 0.25um filters and once in 0.20um strainers (Miltenyi Biotech 130-101-812). Quality control was assayed by measuring live/cell death, cell size and number of clumps. Samples with a viability above 65% were used for sequencing. 10,000 cells per sample were used for sequencing.   A suspension of 10,000 single cells was loaded onto the 10X Genomics Single Cell 3' Chip, and the dowstream cDNA synthesis was performed as per the manufacturer's protocol for the Chromium Single Cell 3' v2 Library (10X Genomics; PN-120233), to generate single cell gel beads in emulsion (GEMs).  Libraries were generated following manufacturer's instructions (ChromiumTM Single Cell 3' Reagent Kits v2).
ENA first public
2025-09-30
INSDC center name
The Francis Crick Institute
INSDC status
public
Submitter Id
E-MTAB-7320_2:Sample 9
broker name
ArrayExpress
collection date
not collected
common name
house mouse
developmental stage
E12.5
genotype
wild type genotype
geographic location (country and/or sea)
not collected
isolate
not applicable
replicate
3
sample name
E-MTAB-7320_2:Sample 9
scientific_name
Mus musculus