characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2022-05-31T00:13:18Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2022-05-31T00:13:18Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA10273798
characteristics.characteristic: External Id
characteristics.text: Department II of Internal Medicine and Center for Molecular Medicine Cologne, University of Cologne
characteristics.characteristic: INSDC center name
characteristics.text: 2022-05-31T00:13:18Z
characteristics.characteristic: INSDC first public
characteristics.text: 2022-05-31T00:13:18Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS7928655
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-11021:Sample 12
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: mIMCD-3
characteristics.characteristic: cell line
characteristics.text: kidney inner medulla collecting duct epithelial cell
characteristics.characteristic: cell type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: Murine inner-medullary collecting duct (mIMCD3, ATCC CRL‐2123) cells were cultured in DMEM-F12 (Sigma-Aldrich) supplemented with 10 % fetal bovine serum (FBS) and 2 mM GlutaMAX (Gibco). For FACS sorted cell lines the medium was additionally supplemented with 100 U/ml Penicillin-Streptomycin (Gibco). mIMCD3 subclone 8 (sc.#8) was selected after FACS sorting the parental line into single cells for a higher genetic homogeneity. For the generation of the CRISPR/Cas9-mediated mIMCD3 sc.#8 mutant cell lines, the cells were seeded in 6-well plates, grown to 50 % confluency and transfected with Lipofectamine 2000 (Invitrogen) according to the manufacturer's instructions. Total RNA extraction was performed with the direct-zol RNA Miniprep kit (Zymo Research) following the manufacture's instructions. Library preparation and sequencing was performed by the Cologne Center for Genomics. Libraries were prepared using the Illumina TruSeq mRNA stranded sample preparation Kit. Library preparation started with 300 ng total RNA. After poly-A selection, mRNA was purified and fragmented using divalent cations under elevated temperature. The RNA fragments underwent reverse transcription using random primers, followed by second strand cDNA synthesis with DNA Polymerase I and RNase H. After end repair and A-tailing, indexing adapters were ligated. The products were then purified and amplified to create the final cDNA libraries. After library validation and quantification (TapStation 4200, Agilent), equimolar amounts of library were pooled and quantified by using the Peqlab KAPA Library Quantification Kit and the Applied Biosystems 7900HT Sequence Detection System.
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: normal
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PATO_0000461
characteristics.characteristic: disease
characteristics.text: Jade2 Knockout (Exon 7)
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: kidney
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UBERON_0002113
characteristics.characteristic: organism part
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: Sample 12
characteristics.characteristic: title