Development of an automated high-throughput DNA extraction protocol using magnetic beads.

摘要

This thesis addresses the efficiency of automated workstations in research laboratories. Chemical reagents used in automated DNA extraction protocols were formulated and manipulated to increase extraction efficiency using magnetic beads as measured by product quality, processing costs, and increased throughput. Additionally, a protocol was tested to establish if multiple steps and reagents could be streamlined to reduce the amount of manipulation involved in sample processing. An evaluation of the Nucleic Acid Tracking System, the Laboratory Information Management System at the NRDPFC, is also performed, which demonstrates the importance of sample tracking for increased throughput. Results establish that the chaotropic buffer formulated to enhance DNA-silica binding reduced costs by 30% when compared to the commercial buffer currently being used. DNA extraction yields were also increased by 20% for tissue samples when compared to the commercial buffer; however, this improvement was not observed with non-invasive low template DNA samples such as scat and hair. Results also demonstrate that the consolidation of current sample lysis reagents into one reagent did not provide an advantage over current sample preparation protocols due to the sample specific requirements relating to each sample type. This thesis illustrates the importance of continually evolving not just the physical aspects of automated workstations, but also perfecting the chemical reagents that are utilized to carry out the extractions in order to decrease costs and increase efficiency to meet the demands of growing laboratories that have increasingly limited resources.;Keywords: DNA extraction, laboratory automation, chaotropic binding buffer, DNA-silica binding, LIMS.