characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2016-03-23T17:02:40Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2016-01-26T12:06:16Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA3856725
characteristics.characteristic: External Id
characteristics.text: Wellcome Trust Sanger Institute, Cambridge, UK EMBL-EBI, Cambridge, UK
characteristics.characteristic: INSDC center name
characteristics.text: 2016-03-23T17:02:40Z
characteristics.characteristic: INSDC first public
characteristics.text: 2016-01-26T12:06:16Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1043859
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-3321:EM_4cell_2_A
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: Embryos were recovered from superovulated F1 (C57Bl6xCBA) mice into M2 media supplemented with 4% BSA, as described previously25. Individual blastomeres were collected at late 2-cell (48 hours after hCG), late 4-cell (10 hours after the first 2-cell blastomere divided), late 8-cell (10 hours after the first 4-cell blastomere divided), 16-cell (78 hours after hCG) and 32-cell (86 hours after hCG) stages. Embryos for blastomere isolation at the 2-, 4,- and 8-cell stages were all collected at the 2-cell stage and cultured in KSOM media (Millipore) at 37C with 5% CO2. Embryos for the 16- and 32-cell blastomeres were collected directly at the respective stages. The zona pellucida of individual embryos was removed using Tyrode's solution (Sigma). Zona-free embryos were incubated for 5 minutes (for the 2,4 and 8-cell stages) or 20 minutes (for the 16 and 32-cell stages) in Ca2+ and Mg2+ free M2 before disaggregation by careful pipetting with a flame-polished glass pipette. Each embryo was processed individually. Single blastomeres for all stages were placed into individual tubes containing 2.3 l of 0.2 % Triton X-100 (Sigma) supplemented with 1 U/l RNAsIN (Ambion). At the late 2-cell stage one blastomere was injected with rhodamine-dextran (Invitrogen), as described previously. Embryos were observed at 20 minute intervals to categorise the order of divisions and whether each cleavage was meridional (M) or equatorial (E), relative to the position of the second polar body. 4-cell stage embryos were sorted into ME, EM, EE and MM groups. Embryos in which the polar body was absent were discarded. Following disaggregation, the individual cells were inspected under an epifluorescent inverted microscope and classified as originating from the first- or second-dividing 2-cell stage blastomere based on the rhodamine fluorescence. Multiplex sequencing libraries were generated from amplified cDNA using Nextera XT (Illumina) and sequenced on a HiSeq 2500 running in rapid mode.
characteristics.characteristic: description
characteristics.text: cleavage 4-cell
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0001285
characteristics.characteristic: developmental stage
characteristics.text: Equatorial - Meridional
characteristics.characteristic: division pattern
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_10090
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: OK
characteristics.characteristic: single cell well quality
characteristics.text: EM_4cell_2_A
characteristics.characteristic: title