Capillary electrophoresis and fluorescence studies on molecular beacon-based variable length oligonucleotide target discrimination
Autor: | Ziad El Rassi, Akhilesh Ramachandran, David W. Goad, Minquan Zhang, Jerry R. Malayer, Glenn A. Olah |
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Rok vydání: | 2003 |
Předmět: |
Conformational change
Fluorophore Clinical Biochemistry Analytical chemistry Biochemistry Analytical Chemistry chemistry.chemical_compound Nucleic acid thermodynamics Capillary electrophoresis Molecular beacon RNA Ribosomal 16S Native state Francisella Fluorescent Dyes Base Sequence Oligonucleotide Temperature Electrophoresis Capillary Nucleic Acid Hybridization Fluorescence RNA Bacterial Spectrometry Fluorescence chemistry Biophysics Nucleic Acid Conformation Oligonucleotide Probes |
Zdroj: | Scopus-Elsevier |
ISSN: | 1522-2683 0173-0835 |
Popis: | Molecular beacons (MBs) are oligonucleotide probes having a compact hairpin structure, with a fluorophore attached to one end and a quencher molecule attached to the other end. In its native state, the fluorophore is quenched by virtue of its proximity to the quencher molecule. Upon hybridization with its complementary oligonucleotide target, fluorescence is elicited due to a conformational change that results in separation of the fluorophore and quencher molecule. The present study describes the hybridization interaction of an MB to various complementary target sequences. The effects of temperature and length of complementary target sequences on hybridization were investigated using capillary electrophoresis and solution-based fluorescence techniques. Hybridization efficiency was dependent on the ability of the target sequences to destabilize the stem region by binding directly to the stem region. Optimal hybridization occurred between 40 and 50 degrees C for all targets tested, with the true target forming a more stable hybrid complex. |
Databáze: | OpenAIRE |
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