Comprehensive Genomic Profiling of BRCA-Associated Cancers
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Corresponding Organization : Ovarian Cancer Action
Other organizations : Cancer Research UK, University of Cambridge, Imperial College Healthcare NHS Trust, University of Turin, Queen Mary University of London, Barts Health NHS Trust, University College Hospital, University College London
Variable analysis
- DNA extraction using QIAmp DNA FFPE Tissue Kit (Qiagen)
- DNA shearing using Covaris LE220 focused-ultrasonicator to produce 100 to 200bp fragments
- Library generation using SureSelect XT standard protocol (Agilent Technologies) for low-input and FFPE samples
- Analysis of PTEN, KRAS, RB1, BRCA2, RAD51B, FANCM, PALB2, RAD51D, TP53, RAD51C, BRIP1, CDK12, NF1, BRCA1, BARD1, and PIK3CA using a custom Ampliseq panel
- Whole-genome sequencing (sWGS) with a minimum of 5 to 10 million reads per sample (mean coverage of 0.1×)
- Nine samples used as a panel of normal controls, five adjacent normal tissue and four whole blood samples
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