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.
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.
