دورية أكاديمية
Multiplexed nucleic acid sensing with single molecule FRET
| العنوان: | Multiplexed nucleic acid sensing with single molecule FRET |
|---|---|
| المؤلفون: | Kaur, Anisa |
| المصدر: | Theses and Dissertations |
| بيانات النشر: | VCU Scholars Compass |
| سنة النشر: | 2021 |
| المجموعة: | Virginia Commonwealth University: VCU Scholars Compass |
| مصطلحات موضوعية: | multiplexing, biosensors, TIRF microscopy, single molecule analysis, nucleic acid sensing, biomarker, Analytical Chemistry |
| الوصف: | In recent years, biomarkers have drawn tremendous interest in the fields of clinical diagnostics and biotechnology. In fact, recent quests for biomarker analyses have discovered a myriad of disease-related biomarkers spanning a wide variety of biomolecules such as proteins, nucleic acids, and small molecule ligands. While the detection of only one target at a time is the most common practice in biosensing today, it has been shown that the accuracy of diagnosis increases significantly by measuring the level of multiple biomarkers instead of just one due to the possible overlap of certain biomarkers in more than one disease. However, currently available ensemble multiplexed methods such as DNA microarrays and polymerase chain reactions (PCR) often suffer from complicated design/engineering, semi-quantitative analysis, or a high false positive rate. Single molecule techniques can be an asset, in this regard, by providing unique insights into the behavior and interactions of individual molecules. For example, single molecule fluorescence resonance energy transfer (smFRET) is able to quantitatively measure signals from individual molecules and is a sensitive means for detection of biomarkers by relying on the emission from one or more donor/acceptor FRET pairs. However, current methods require multiple excitation sources and complex data analysis algorithms. To address this knowledge gap, using prism-based total internal reflection fluorescence (pTIRF) microscopy, we developed a FRET-based multiplexed detection method for DNA sequences utilizing only a single donor/acceptor fluorophores pair. By tuning the minimum distance separation between a FRET pair through a careful design, we showed that multiple FRET values can be achieved depending on the distance set. We utilized interconvertible hairpin-based sensors (iHabSs) where detection was dependent on a toehold-mediated displacement of a probe strand by the target. This approach allowed for simultaneous detection of 3 different DNA sequences with a detection limit of ... |
| نوع الوثيقة: | text |
| وصف الملف: | application/pdf |
| اللغة: | unknown |
| العلاقة: | https://scholarscompass.vcu.edu/etd/6644Test; https://scholarscompass.vcu.edu/cgi/viewcontent.cgi?article=7781&context=etdTest |
| الإتاحة: | https://scholarscompass.vcu.edu/etd/6644Test https://scholarscompass.vcu.edu/cgi/viewcontent.cgi?article=7781&context=etdTest |
| حقوق: | © The Author |
| رقم الانضمام: | edsbas.7B445BFC |
| قاعدة البيانات: | BASE |
| الوصف غير متاح. |