characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2017-09-20T17:02:02Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2017-09-18T17:37:25Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA104306092
characteristics.characteristic: External Id
characteristics.text: BRISTOL
characteristics.characteristic: INSDC center name
characteristics.text: 2017-09-20T17:02:02Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-09-18T17:37:25Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1931070
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-6071:Sample 2
characteristics.characteristic: Submitter Id
characteristics.text: 7
characteristics.unit: day
characteristics.characteristic: age
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: Targeting of the Bhlhe23 gene and genotyping of the Bhlhe23 allele was previously described by Kim et al 2008. An approximately 5-kb HindIII/NotI DNA fragment encompassing the Bhlhb4 gene was isolated by PCR from 129/Sv genomic DNA. A loxP-flanked PGK-neomycin resistance cassette in reverse orientation was introduced into the KpnI site approximately 1 kb upstream of the Bhlhb4 gene. A third loxP sequence was introduced approximately 300 bases distal to the Bhlhb4 3′ UTR, between adjacent BsrGI and MunI restriction sites. A PGK-diptheria toxin gene cassette was introduced at the 3′ end of the Bhlhb4 locus. This vector was electroporated into mouse 129 J1 embryonic stem (ES) cells. A null allele was generated by transiently transfecting targeted ES cells with a Cre recombinase plasmid, pOG231. ES cells carrying the deletion allele were microinjected into C57BL/6 blastocysts, and chimeras were tested for germline transmission of the mutant allele by breeding to C57BL/6 females and PCR genotyping of pups. PN7 wildtype and mutant eyes were removed following termination of the animals. Eyes were dissected in ice cold PBS. The cornea was pierced and the eye pulled apart. The retina was then gently pulled away from the rest of the eye. Retinal samples were stored in RNAlater® (Qiagen) at -20C. RNA extraction and purification was performed using the RNeasy® Mini kit from Qiagen (Hildern, Germany) before quantification on a Nanodrop spectrophotometer (ND-1000, Thermo Fisher Scientific). Barcoded sequencing libraries were prepared using the Illumina® TruSeq Stranded Total RNA Low-Throughput sample RiboZero Human/Rat/Mouse prep kit following manufacturer’s guidelines (Illumina, San Diego, CA, USA). Libraries were quantified with a Qubit® fluorometer (Thermo Fisher Scientific) using the double stranded DNA High sensitivity assay.
characteristics.characteristic: description
characteristics.text: normal
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PATO_0000461
characteristics.characteristic: disease
characteristics.text: wild type genotype
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0005168
characteristics.characteristic: genotype
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: retina
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UBERON_0000966
characteristics.characteristic: organism part
characteristics.text: litter 2
characteristics.characteristic: replicate
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: male
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PATO_0000384
characteristics.characteristic: sex
characteristics.text: C56BL/6
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0004472
characteristics.characteristic: strain
characteristics.text: Sample 2
characteristics.characteristic: title