Large -scale targeted NGS sequencing made easy & a ffordable. BRCA Gene Panel Preface: The most common of all cancers to affect women is breast cancer, with as many as one-in-eight women contracting it during their lifetime. Breast cancer is a complex disease with many causes, however we do know that approximately 10% of all cases are due to an individual’s DNA mutation. These hereditary mutations commonly result in the early-onset of breast cancer, recurrent breast cancer and breast cancer involving both breasts. Approximately 25-50% of hereditary breast and ovarian cancer cases can be explained by mutations in the BRCA1 and BRCA2 genes. A comprehensive hereditary breast and ovarian cancer risk assessment should include germline sequencing of the BRCA1 and BRCA2 genes as well as additional key genes with a known association of breast/ovarian cancer. Individuals with a suspected inherited genetic cancer risk detected in the Breast Cancer Panel will have a clinical advantage to affect patient medical management, guide preventative measures, direct surgical options and estimate personal and familial cancer risk. Methodology: The Otogenetics Breast Cancer Panel is performed by next generation sequencing using custom oligonucleotide-based target capture followed by Illumina HiSeq sequencing of the coding regions of the Breast Cancer panel genes; with >100 fold of average coverage and no gap in the capture of targeted genes. Clinically significant variants are identified and reported. Results: Each Otogenetics Breast Cancer Panel report includes a detailed explanation of all clinically-relevant variants. Breast Cancer gene panel & other disease panel reagent kits also available. Visit www.otogenetics.com to learn more. The Otogenetics Breast Cancer Panel allows the clinician or researcher to sequence 19 genes known to be high-risk breast cancer susceptibility genes (see back for gene list). Testing Rationale: The Otogenetics Breast Cancer Panel allows the identification of the specific risk or cause factor for breast cancer in the patient, thus aid in eliminating unnecessary testing. This is an important part of a comprehensive cancer risk evaluation, and identification of a specific genetic mutation provides important information for at-risk patients, including those with the following:     Two primary breast cancers or clustering of breast and ovarian cancer. Early-onset breast cancer (<50 years-of-age) or bilateral breast cancer. Presence of male breast cancer. Ovarian cancer at any age. Test Name: BRCA Turn-Around-Time: Approximately 5-6 weeks Specimen Requirement: 5 mL whole blood, or by saliva collection kit. 400 Pinnacle Way, Ste 435, Norcross, GA 30071 USA • [email protected] • +1-855-686-4363 • www.otogenetics.com BRCA Gene Panel OMIM NCBI Reference (Online Mendelian Inheritance in Man) CDS bps # of Exons AR NM_000044.3 313700 Androgen receptor 10661 8 ATM NM_000051.3 607585 Serine-protein kinase ataxia telangiectasia mutated 13147 68 BARD1 NM_000465.3 601593 BRCA associated RING domain 1 5523 11 BRCA1 NM_007298.3 113705 Breast cancer 1, early onset 3699 22 BRCA2 NM_000059.3 600185 Breast cancer 2, early onset 11386 27 BRIP1 NM_032043.2 605882 BRCA1-interacting protein 8166 20 CASP8 NM_001080124.1 601763 Apoptosis-related cysteine protease 8 2750 9 CDH1 NM_004360.3 192090 Cadherin 1 4815 16 CHEK2 NM_001005735.1 604373 Serine/threonine checkpoint kinase 2 1991 16 DIRAS3 NM_004675.2 605193 GTP-binding Ras-like protein 3 1642 2 ERBB2 NM_001005862.1 164870 Avian erythroblastic leukemia viral oncogene homolog 2 4816 30 NBN NM_002485.4 602667 Nibrin 4639 16 PALB2 NM_024675.3 601355 Partner and localizer of BRCA2 4069 13 PTEN NM_000314.4 601728 Phosphatase and tensin 5572 9 RAD50 NM_005732.3 604040 DNA repair protein RAD50 homolog 6597 25 RAD51 NM_001164269.1 179617 DNA repair protein RAD51A homolog 2147 10 STK11 NM_000455.4 602216 Serine/threonine protein kinase 11 3286 10 TGFB1 NM_000660.4 190180 Transforming growth factor β1 2217 7 TP53 NM_000546.5 191170 Tumor protein p53 2591 11 Gene Description 2 hours 2 hours 3 days 2.5 hours 2 hours 2 hours  Proceed to Sequencing via Illumina HiSeq/MiSeq  Assess Enrichment Using qPCR (Oto-BRCA-Primer)  Amplify Captured DNA (Oto-Enrich-001)  Wash and Recover Captured DNA (Oto-Enrich-001)  Hybridize Samples to Oto-BrCa-Probe (Oto-BRCA-Probe) 4 hours  Pooling of Amplified Multiplex DNA Libraries TIME  DNA Library Amplification Using LM-PCR (Oto-DLP-001) STEP DNA Fragmentation, End Repair, Tailing, Ligation (Oto-DLP-001) TotSeq™ Targeted Human BRCA Genes Sample Preparation Flowchart 27 hours or 10 days Copyright 2014 by Otogenetics Corporation. Products and specifications are subject to review and change without notice. 060914 For Research Use Only 400 Pinnacle Way, Ste 435, Norcross, GA 30071 USA • [email protected] • +1-855-686-4363 • www.otogenetics.com