Homogenous M13 bacteriophage quantification assay using switchable lanthanide fluorescence probes
| Autor: | Tero Soukka, Urpo Lamminmäki, Ari Lehmusvuori, Tuomas Huovinen, Julius Manninen |
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| Rok vydání: | 2012 |
| Předmět: |
Lanthanide
M13 bacteriophage Models Genetic biology ta1183 DNA Single-Stranded Nucleic Acid Hybridization biology.organism_classification Lanthanoid Series Elements Combinatorial chemistry Fluorescence Molecular biology General Biochemistry Genetics and Molecular Biology Complementation Bacteriophage chemistry.chemical_compound chemistry Limit of Detection ta318 Oligonucleotide Probes DNA Bacteriophage M13 Fluorescent Dyes Single-Chain Antibodies Biotechnology |
| Zdroj: | BioTechniques. 53:301-303 |
| ISSN: | 1940-9818 0736-6205 |
| DOI: | 10.2144/0000113954 |
| Popis: | We have developed a rapid and reliable bacteriophage quantification method based on measurement of phage single-stranded DNA (ssDNA) using switchable lanthanide chelate complementation probes. One oligonucleotide probe contains a non-fluorescent lanthanide ion carrier chelate and another probe is labeled with a light absorbing antenna ligand. Hybridization of the non-fluorescent complementation probes in adjacent positions on the released bacteriophage ssDNA leads to high local concentrations of the lanthanide ion carrier chelate and the antenna ligand, inducing formation of a fluorescent lanthanide chelate complex. This method enables monitoring of bacteriophage titers in a 20 min assay with a dynamic range of 109–1012 cfu/mL in a microtiter well format. While designed for titering filamentous bacteriophage used in phage display, our method also could be implemented in virological research as a tool to analyze ssDNA virus reproduction. |
| Databáze: | OpenAIRE |
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