National Epidemiologic Survey on Alcohol and Related Conditions: DNA Repository

描述（由申请人提供）：全国酒精及相关疾病流行病学调查（NESARC I和II）是美国平民非机构人口中规模最大、最雄心勃勃的酒精中毒及相关疾病流行病学调查。NESARC-III更加雄心勃勃，它的目标是收集多达46,500名受试者的唾液样本，并从中提取DNA，通过大规模DNA重测序来确定酒精使用障碍（AUDs）和共病障碍的遗传风险因素，并检查基因与环境的相互作用。我们建议为NIAAA NESARC-III研究建立唾液DNA库。我们的目标是：1；从人类唾液样本中提取基因组DNA：我们将采用最先进的自动化技术和基于珠状化学来富集高分子量的人类基因组DNA，这些DNA相对没有污染物，也没有RNA和不需要的DNA物种（即细菌DNA污染）。2. 全面的核酸质量控制：我们设计并验证了一种新的，全面的分析和功能质量控制（QC）程序，确保DNA的最高质量及其对未来测序分析的效用。分析QC包括吸光度和染料浓度和纯度测量，精确的非接触式体积测量，以及每个样品中人类基因组DNA量的特定定量。功能QC采用高通量，分子遗传指纹SNP测定（RUID）。我们还将使用几个不同的下一代测序平台来演示这种唾液DNA在未来测序中的效用。3. 集成的核酸存储和分配：所有NESARC-III样品将存储在专用冷冻室中，温度为-80摄氏度，并在安全的设施中进行全天候监控，该设施包含最先进的冗余电源和HVAC系统。所有样品提取、质量控制、储存、生物材料请求和分发工作流程都由RUCDR STARLIMS（一个企业级实验室信息管理系统）记录和管理。

DESCRIPTION (provided by applicant): The National Epidemiological Surveys on Alcohol and Related Conditions (NESARC I and II) are the largest and most ambitious epidemiological surveys on alcoholism and related disorders in the civilian, non-institutionalized US population. NESARC-III is even more ambitious as it aims to collect saliva samples from as many as 46,500 subjects and extract DNA from each in an effort to identify genetic risk factors for alcohol use disorders (AUDs) and co-morbid disorders through large-scale DNA re-sequencing and to examine gene-environment interactions. We propose to establish the Saliva DNA Repository for the NIAAA NESARC-III study. Our Aims are: 1. Genomic DNA Extraction from Human Saliva Samples: We will employ state-of-the-art automation and bead-based chemistry to enrich for high molecular weight human genomic DNA relatively free of contaminants and of RNA and undesirable DNA species (i.e., bacterial DNA contamination). 2. Comprehensive Nucleic Acid Quality Control: We have engineered and validated a novel, comprehensive analytical and functional quality control (QC) program that ensures the highest quality of DNA and its utility for future sequencing analyses. Analytical QC consists of absorbance and dye-based concentration and purity measurements, precision non-contact volume measurements, and specific quantification of the amount of human genomic DNA in each sample. Functional QC employs a high-throughput, molecular genetic fingerprinting SNP assay (the RUID). We will also use several different Next Generation sequencing platforms to demonstrate the utility of this saliva DNA for future sequencing. 3. Integrated Nucleic Acid Storage and Distribution: All NESARC-III samples will be stored at -80 degrees C in dedicated freezers that are monitored 24/7/365 in a secure facility that contains state-of-the-art redundant power and HVAC systems. All sample extraction, QC, storage, biomaterial requests and distribution workflows are documented and managed by the RUCDR STARLIMS, an enterprise-level laboratory information management system. PUBLIC HEALTH RELEVANCE: Alcohol use disorders (AUDs) cost our nation billions of dollars due to lost productivity, provision of medical care, accidents, violence and the breakup of social order. In addition, they are responsible for much human misery and suffering. We must understand the causes, onset and lifetime progression of AUDs through epidemiology and genetics studies if we are to develop better treatment and prevention strategies.