Ultrahigh speed Direct PCR. A method for obtaining STR based genotypes in under 6 minutes

As submitted by the proposer: There are situations in which it is very important to rapidly screen crime scene samples and unknown individuals who may have been involved in a crime. Examples include seized evidence potentially linked to a suspect, co-mingled bone samples buried in a mass grave, or the determination of which blood stains present at a crime scene may be probative. In these situations many samples may need to be run in order to create probable cause for detaining a suspect or sorting through excessive amounts of evidence. Current DNA typing methods provide the best biometric information yielding identity, kinship and geographical origin, but they are not sufficiently fast to permit identity of a suspect DNA in real time. Current rapid DNA systems take about 90 minutes and laboratory DNA analysis is even slower. Direct PCR coupled with high speed polymerases can greatly speed the processing time from sample to STR genotype. Coupling these advances with new thermal cycler designs can make the analysis even faster. Using a flow based thermal cycler from Thermal Gradient, we can now perform STR amplifications in 4 minutes and with our modified Agilent Bioanalyzer that can produce a genotype in 80 seconds. This means we will have the capability to produce genotypes in under 6 minutes. It is our goal to develop a 7 locus CODIS based genotype for quick, highly informative sample screening process that also retains sufficient DNA for later manual processing of the full genotype. With such a procedure in place a crime lab can produce a nearly instantaneous genotype from buccal samples and blood stains. This new technology should open all kinds of new applications for direct PCR, with analysis speeds previously only attainable on science fictions shows like Star Trek. The project will involve a collaborative effort between the FIU Department of Chemistry, Steve Lee from the Department of Justice Studies at San Jose State University and the Broward Sheriff’s Office DNA Laboratory. Our industrial partners will include DNA Polymerase Technology (supplier of polymerases), Thermal Gradient (supplier of high speed thermal cycling) and Agilent Technologies (developer of high speed microfluidic DNA chips) This project is interdisciplinary involving the efforts of academia, instrument manufacturers, biochemistry research and law enforcement. This project contains a research and/or development component, as defined in applicable law. ca/ncf