Single-Molecule Fluorescence Using Nucleotide Analogs: A Proof-of-Principle
| Autor: | Karin Musier-Forsyth, Chamaree de Silva, David Rueda, Eric M Patrick, Elvin A. Alemán |
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| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
DYNAMICS
Technology CHEMISTRY PHYSICAL NANOSCIENCE & NANOTECHNOLOGY Biophysical Chemistry and Biomolecules Materials Science PHYSICS ATOMIC MOLECULAR & CHEMICAL 2-Aminopurine MATERIALS SCIENCE MULTIDISCIPLINARY Biology BASE STACKING chemistry.chemical_compound DINUCLEOTIDES EXCITATION General Materials Science Nucleotide PSPGI Physical and Theoretical Chemistry PROBE 2-AMINOPURINE FLUORESCENCE chemistry.chemical_classification Science & Technology 02 Physical Sciences Physics RNA DNA Single-molecule experiment Fluorescence Combinatorial chemistry 3. Good health Chemistry chemistry Physical Sciences Click chemistry Nucleic acid Science & Technology - Other Topics 03 Chemical Sciences ABASIC SITES |
| Zdroj: | The Journal of Physical Chemistry Letters |
| Popis: | Fluorescent nucleotide analogues, such as 2-aminopurine (2AP) and pyrrolo-C (PyC), have been extensively used to study nucleic acid local conformational dynamics in bulk experiments. Here we present a proof-of-principle approach using 2AP and PyC fluorescence at the single-molecule level. Our data show that ssDNA, dsDNA, or RNA containing both 2AP and PyC can be monitored using single-molecule fluorescence and a click chemistry immobilization method. We demonstrate that this approach can be used to monitor DNA and RNA in real time. This is the first reported assay using fluorescent nucleotide analogs at the single-molecule level. We anticipate that single 2AP or PyC fluorescence will have numerous applications in studies of DNA and RNA, including protein-induced base-flipping dynamics in protein–nucleic acid complexes. |
| Databáze: | OpenAIRE |
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