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dc.contributor.authorIacona, Joseph Robert, Jr.en_US
dc.date.accessioned2017-04-13T01:28:04Z
dc.date.issued2013
dc.date.submitted2013
dc.identifier.urihttps://hdl.handle.net/2144/21180
dc.descriptionThesis (M.S.F.S.) PLEASE NOTE: Boston University Libraries did not receive an Authorization To Manage form for this thesis or dissertation. It is therefore not openly accessible, though it may be available by request. If you are the author or principal advisor of this work and would like to request open access for it, please contact us at open-help@bu.edu. Thank you.en_US
dc.description.abstractBiological evidence may contain any number of cells in any proportion. Extreme low-template DNA samples are often very difficult to interpret due to complex signal or peaks which may be indistinguishable from baseline noise. Current solutions focus on increasing the amount of amplicon detected by adjusting PCR cycle number or capillary electrophoresis injection parameters. Consensus profiling is an additional option. However, the aforementioned solutions are often not helpful for extreme low-template samples due to the high occurrence of allelic drop-out. Additionally, PCR is a destructive technique that causes one amplification to completely exhaust this type of sample, making further typing and analysis impossible. Therefore, a technique that allows for the re-generation of a DNA template in order to amplify it multiple times would be an extremely useful tool. This study outlines the development of a method that allows for the recursive amplification of a DNA sample. Amplification was performed using biotinylated primers for an STR locus and the resulting product was cleaned using streptavidin-coated magnetic beads to sequester the amplicons. Subsequent centrifugal filtration was used to remove the remaining PCR components, thus isolating the original genomic DNA. Re-amplification was then successfully performed at a different STR locus. Though successful, multiple run-throughs of the method indicated retention of signal from the original amplification as well as significant genomic DNA loss during the process. This study outlines experiments seeking to characterize the cause(s) of these imperfections in order to effectively direct method optimization. A computer generated dynamic model was also created and used to simulate the recursive amplification process to assist in development. When optimized, it is expected that recursive amplification can significantly reduce the difficulties associated with low-template DNA analysis and eradicate the concept of an ‘exhaustive’ DNA sample.en_US
dc.language.isoen_US
dc.publisherBoston Universityen_US
dc.subjectForensic scienceen_US
dc.subjectBioforensicsen_US
dc.subjectRecursive genotypingen_US
dc.subjectGenomic DNA isolationen_US
dc.titleGenomic DNA isolation from amplified product for recursive genotyping of low-template DNA samplesen_US
dc.typeThesis/Dissertationen_US
dc.description.embargo2031-01-01
etd.degree.nameMaster of Science in Forensic Scienceen_US
etd.degree.levelmastersen_US
etd.degree.disciplineBioforensicsen_US
etd.degree.grantorBoston Universityen_US


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