characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2015-03-19T17:02:31Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T21:11:25Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2781633
characteristics.characteristic: External Id
characteristics.text: Faculty of Science, University of Zagreb
characteristics.characteristic: INSDC center name
characteristics.text: 2015-03-19T17:02:31Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T21:11:25Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS551380
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-2950:s_Blast.WE
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: Growing oocytes were obtained from 12-14-day-old B6D2F1 mice (SLC Japan). The ovaries were transferred to HEPES-buffered KSOM and punctured with a 30-gauge needle. Liberated oocytes with attached follicle cells were harvested and the follicle cells were gently removed using a narrow-bore glass pipette. The oocytes were then transferred into alpha-minimal essential medium (alpha-MEM; Life Technologies, Inc., Grand Island, NY) containing 5% fetal bovine serum and 10 ng/ml epidermal growth factor (both from Sigma-Aldrich). Total RNA was extracted from 3000, 3000, 4500, 2800, 1400 and 700 MII eggs, 1-, 2-, and 4-cell embryos, morulae and blastocysts obtained from C57BL6/J mice, respectively, using Isogen (Nippon Gene, Tokyo, Japan), according to the manufacturers instructions. In the second trials for MII eggs and 1-cell embryos, similar numbers of cells used for the first trials were used for RNA extraction. RNA quality analysis and size-selection (>200 tn) were performed on the Bioanalyzer RNA Pico Chip (Agilent Technologies, Santa Clara, CA). RNA-Seq libraries were constructed using the mRNA-Seq Sample Preparation Kit (Illumina, San Diego, CA) without selection of polyadenylated RNA. Briefly, size-selected total RNA was fragmented to 40~900 nt fragments in fragmentation buffer at 94 oC for 5 min, reverse transcribed with random primers and ligated with adaptors. cDNA templates were amplified by PCR in the conditions as follows: the initial heat treatment at 98oC for 30 s followed by 15 cycles of 98oC for 10 s, 65oC for 30 s, 72oC for 30 s, and final treatment of 72 oC for 5 min. RNA-seq libraries were subjected to 76-nt paired-end-sequencing.
characteristics.characteristic: description
characteristics.text: blastocyst
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UBERON_0000358
characteristics.characteristic: developmental stage
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: C57BL6/J
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0000606
characteristics.characteristic: strain
characteristics.text: s_Blast.WE
characteristics.characteristic: title