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Using PNA Probes for Hybridization-Based Analysis of miRNAs in Capillary Electrophoresis

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dc.contributor.advisorKrylov, Sergey N.
dc.contributor.authorAnand, Mansi
dc.date.accessioned2019-11-22T18:36:33Z
dc.date.available2019-11-22T18:36:33Z
dc.date.copyright2019-04
dc.date.issued2019-11-22
dc.date.updated2019-11-22T18:36:32Z
dc.degree.disciplineChemistry
dc.degree.levelMaster's
dc.degree.nameMSc - Master of Science
dc.description.abstractThe inherent separation inefficiencies of the excess probes and hybrids and the hybrids from each other limits the use of capillary electrophoresis (CE) for hybridization-based analysis of microRNA (miRNAs). Our lab developed direct quantitative analysis of multiple miRNA (DQAMmiR) to address the above using two types of mobility shifters: single-strand DNA binding protein (SSB) added to the CE run buffer and peptide drag tags conjugated with the probes. Here, we introduce second-generation DQAMmiR, which is simpler than the first-generation as it omits the addition of SSB protein in CE run buffer by the use of uncharged Peptide Nucleic Acid (PNA) instead of DNA probes. Additionally, the performance of the assay was also validated in the presence of prostate cancer (PCa)-derived crude cell lysate for two deregulated miRNAs. With ongoing improvements, PNA-facilitated DQAMmiR holds great potential for reliable direct quantitative analysis of endogenous miRNAs in clinical samples.
dc.identifier.urihttp://hdl.handle.net/10315/36664
dc.languageen
dc.rightsAuthor owns copyright, except where explicitly noted. Please contact the author directly with licensing requests.
dc.subjectAnalytical chemistry
dc.subject.keywordsAnalytical Chemistry
dc.subject.keywordsCapillary electrophoresis
dc.subject.keywordsPeptide nucleic acid (PNA)
dc.subject.keywordsmicroRNA (miRNA) analysis.
dc.titleUsing PNA Probes for Hybridization-Based Analysis of miRNAs in Capillary Electrophoresis
dc.typeElectronic Thesis or Dissertation

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