High Accuracy Genotyping of Complex Mixtures and Damaged DNA
Technical Report,21 Sep 2015,20 Sep 2018
University of Washington Seattle United States
Pagination or Media Count:
Major Goals Objective 1 Define and validate a standardized set of equipment and reagents and provide a step by-step protocol that can be performed by a laboratory technician. This objective will entail developing quality control QC methods to monitor the success of the enzymatic steps required for sample processing in order to ensure consistent performance and maximize the success rate of genotyping difficult samples. Accuracy, precision, sensitivity, and specificity will be evaluated. Objective 2 Provide a basic data analysis workflow for genotype calling. This objective will create a computational workflow for genotype calling and forensic identification from DS analyses of heterogeneous mixtures of low quality DNA. The workflow will have two operational modes 1 If a target genotype is provided, then the workflow will determine if a particular individual or individuals relative DNA is present. 2 If no specific target genotype is provided, then the workflow will produce a list of possible genotypes present. Objective 3 Validate protocol and equipment for use in genotyping complex DNA mixtures and highly damageddegraded DNA. This objective will use DS to evaluate DNA samples subjected to a number of environmental conditions, such as increasing mixture complexity, extended exposure to UVsunlight, elevated temperatures, and damaging chemicals. In this way, the limits of DS to genotype highly complex and damaged DNA will be defined. Objective 4 Develop and provide a training program and material. Once DS has been optimized and validated for forensic analysis, we will develop and provide training and hands on experience for technical staff on how the system functions and how sample preparation is performed.