Validation and Optimization of Automated Instrumentation for Mitochondrial DNA Database Sample Processing

Crispin, Shelley Jeanine. Validation and Optimization of Automated Instrumentation for Mitochondrial DNA Database Sample Processing. Master of Science (Forensic Genetics), August 2004, 43 pages, 8 figures, 8 tables, 26 references. The validation of Qiagen 9604 BioRobot for DNA extraction and two Corbett CAS-1200 Liquid Precision Handling Systems for mitochondrial DNA database sample processing will alleviate sample backlogs and increase sample throughput. A manual cross-contamination study and an automated cross-contamination study showed that the use of automated instrumentation allowed for quicker and more accurate sample processing. The validation process was completed by processing a 96-well high throughput plate containing AFDIL employee samples. AFDIL loses approximately 10% of samples cycle sequenced using quarter volume BigDye terminator cycle reactions. To validate the use of half volume BigDye terminator cycled sequencing a 96-well high throughput plate containing AFDIL employee samples was used to compare the percentage of samples lost. On average 1.6% of the samples were lost per 96-well high throughput plate utilizing half volume BigDye terminator cycle sequencing reactions. This study has shown that the validation of half volume BigDye terminator cycle sequencing reactions will minimize the number of samples that will have to be reprocessed per 96-well high throughput plate.