characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2022-09-22T00:41:13Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2022-09-22T00:41:13Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA110424403
characteristics.characteristic: External Id
characteristics.text: LUMC
characteristics.characteristic: INSDC center name
characteristics.text: 2022-09-22T00:41:13Z
characteristics.characteristic: INSDC first public
characteristics.text: 2022-09-22T00:41:13Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS12522767
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-11971:SV-FLB_6_KCNH2_Acc_A561T
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: LUMCi028-A
characteristics.characteristic: cell_line
characteristics.text: induced pluripotent stem cell
characteristics.characteristic: cell_type
characteristics.text: human
characteristics.characteristic: common name
characteristics.text: Protocols: Approximately 1x10^6 hiPSCs (cell line described here: https://hpscreg.eu/cell-line/LUMCi028-A) were rinsed once with PBS without Mg2+ and Ca2+ (PBS-/-) and dissociated to single cells with 1x TrypLE Select (ThermoFisher). The dissociation reagent was neutralized with culture medium (DMEM/F12, BSA 0.5%) and the pellet of cells was resuspended in PBS-/- and kept at -20°C until the genomic DNA extraction protocol was performed. Genomic DNA was extracted from hiPSCs using the High Pure PCR Template Preparation Kit (Roche) and treated with RNAse A (10 mg/ml, Invitrogen) according to the manufacturers’ instructions. 500 ng of gDNA was fragmented by sonication, followed by Illumina library prep using the KAPA Hyper Kit and unique barcoding of each sample with IDT adapters (xGen UDI-UMI adapters). The samples were purified twice to remove any adapter dimers and fragments below 150bp. The adapter ligated samples were further PCR enriched using the p5 and p7 primers for 10x cycles using the KAPA HiFi mastermix.The Hyb Module box of the SureSelectQXT Target Enrichment for the Illumina Platform (Agilent) was used for hybridization and capturing of each sample separately. Briefly, 750 ng of the Illumina-prepped sample and 500ng of the biotinylated RNA capture probes were used in the reaction. A 15 cycle post capture PCR with the p5 and p7 primers was performed using the KAPA HiFi mastermix, with the resulting samples purified using 0.8x Ampure XP beads (Beckman Coulthier) and quantified using the Qubit 2.0 (ThermoFisher).
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: Familial long QT syndrome
characteristics.characteristic: disease
characteristics.text: NM_000238.4:c.1681G>A
characteristics.characteristic: genetic modification
characteristics.text: genetically modified genotype
characteristics.characteristic: genotype
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: skin
characteristics.characteristic: organism part
characteristics.text: fibroblast
characteristics.characteristic: progenitor cell type
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: SV-FLB_6_KCNH2_Acc_A561T
characteristics.characteristic: title