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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Degenerate Oligonucleotide Primed-PCR: Thermalcycling Modifications and Comparison Studies

Rodier, Denise N. 01 January 2006 (has links)
Degenerate Oligonucleotide Primed-PCR (DOP-PCR) can potentially enhance analysis of low copy number DNA samples. Theoretically, this procedure replicates fragments of the genome that can then be used for downstream multiplex STR analysis. The objective of this study is to optimize DOP-PCR by examining ramplelongation times and cycle numbers in the non-specific amplification portion of DOP-PCR, and by modifying the degenerate primer. Additionally, other methods such as Multiple Displacement Amplification (MDA) and Low Copy Number PCR (LCN PCR) were examined for their ability to create accurate DNA profiles from low DNA input amounts. Increasing the ramplelongation times showed no effect on downstream STR amplification success. An increase of cycle number increased DNA yield, but STR amplification success was undetermined. Although modifying the degenerate primer to one with a higher degeneracy decreased DNA yield, it ultimately improved STR amplification success. In comparison studies, LCN PCR produced higher STR amplification success than MDA.

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