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dc.contributor.authorCoe, Genevieve
dc.date.accessioned2015-10-07T20:37:35Z
dc.date.available2015-10-07T20:37:35Z
dc.date.issued2015-05
dc.identifier.urihttp://hdl.handle.net/10675.2/579480
dc.descriptionThis file is restricted to Augusta University. Please log in using your JagNet ID and password to access.en_US
dc.description.abstractAptamers are composed of oligonucleotides and are most recognized for their ability to function much like antibodies. Unlike antibodies, aptamers can be generated in vitro, have a longer shelf life and are potentially faster to synthesize. The current method of aptamer generation is through a cycle known as SELEX, which stands for Systematic Evolution of Ligands through Exponential Enrichment. The SELEX method traditionally uses a library of single-stranded DNA or RNA as a starting material, which is incubated with a target. The nucleotides that bind to the target are separated from the unbound nucleotides, eluted from the target and amplified by PCR. The amplified nucleotides go through several more cycles of SELEX until aptamers with a high binding affinity for the target are produced. Before the double-stranded products from PCR can be used in another cycle they have to be separated into single-strands. The process of regenerating single-stranded DNA from double-stranded DNA is often time consuming and results in a low product yield, decreasing the efficiency of the SELEX process. In order to improve the efficiency of aptamer generation, we explored the use of a novel starting material based on its potential to make SELEX a more automated process. Stem-loop DNA has both double-stranded and single-stranded DNA portions due to its unique secondary structure. It has the ability to retain the single-stranded portion of its special conformation after PCR amplification. After characterization of a specifically designed stem-loop, we incorporated the use of stem-loop DNA as a starting material in SELEX to potentially bypass the regeneration of single-strand DNA in SELEX. The success of this modified approach to SELEX could result in a more efficient method for the generation of aptamers.en_US
dc.language.isoen_USen_US
dc.publisherAugusta Universityen_US
dc.rightsCopyright protected. Unauthorized reproduction or use beyond the exceptions granted by the Fair Use clause of U.S. Copyright law may violate federal law.en_US
dc.subjectSELEXen_US
dc.subjectDNA, Single-Strandeden_US
dc.subjectRNAen_US
dc.titleNovel Use of Stem-loop DNA in SELEXen_US
dc.typeThesisen_US
dc.contributor.departmentDepartment of Chemistry and Physicsen_US
dc.description.advisorSpencer, Angie


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