Day 7 Embryo 7
Homo sapiens
Protocols: Embryos were placed in drops of G-MOPS solution (Vitrolife; Cat. No. 10129) on a petri dish overlaid with mineral oil. The plate was placed on a microscope stage (Olympus IX70) and the embryos were held with an opposing holding pipette and blastomere biopsy pipette (Research Instruments) using micromanipulators (Narishige, Japan). The biopsy mode of a Saturn 5 laser (Research Instruments) was used to separate the mural TE from the ICM and polar TE. The separated mural TE was transferred to individual low bind RNAse-free tube containing 0.25 μl RNase inhibitor, 4.75 μl Dilution buffer (SMARTer Ultra Low Input RNA kit, Clontech; Cat. No. 634820) and 5 μl nuclease-free water on a pre-chilled CoolRack (Biocision, CA). Samples were stored at - 80°C until ready to be processed. cDNA was synthesized using SMARTer Ultra Low Input RNA for Illumina Sequencing-HV kit (Clontech Laboratories; Cat. No. 634820) according to the manufacturer's guidelines and as previously published (Blakeley et al., 2015b; Hyslop et al., 2016). cDNA was sheared using Covaris S2 with the modified settings 10% duty, intensity 5, burst cycle 200 for 2 min. Libraries were prepared using Low Input Library Prep Kit (Clontech Laboratories; Cat. No. 634900) according to the manufacturer's instructions. Library quality was assessed with an Agilent 2100 BioAnalyser and concentration measured by QuBit broad range assay (Thermo Fisher; Cat. No. Q32850).
ENA-FIRST-PUBLIC
2023-07-19T00:29:15Z
ENA-LAST-UPDATE
2023-07-19T00:29:15Z
External Id
SAMEA9424065
INSDC center alias
KING'S COLLEGE LONDON
INSDC center name
KING'S COLLEGE LONDON
INSDC first public
2023-07-19T00:29:15Z
INSDC last update
2023-07-19T00:29:15Z
INSDC status
public
Submitter Id
E-MTAB-10749:Day 7 Embryo 7
age
7
broker name
ArrayExpress
common name
human
developmental stage
embryo stage
individual
Embryo 7
isolate
not applicable
organism part
blastocyst
sample name
E-MTAB-10749:Day 7 Embryo 7
scientific_name
Homo sapiens