Homogenous M13 bacteriophage quantification assay using switchable lanthanide fluorescence probes
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 la...
| Veröffentlicht in: | BioTechniques. - 1988. - 53(2012), 5 vom: 13. Nov., Seite 301-3 |
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| Weitere Verfasser: | , , , |
| Format: | Online-Aufsatz |
| Sprache: | English |
| Veröffentlicht: |
2012
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| Zugriff auf das übergeordnete Werk: | BioTechniques |
| Schlagworte: | Journal Article Research Support, Non-U.S. Gov't DNA, Single-Stranded Fluorescent Dyes Lanthanoid Series Elements Oligonucleotide Probes Single-Chain Antibodies |
| Zusammenfassung: | 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 10(9)-10(12) 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 |
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| Beschreibung: | Date Completed 19.03.2013 Date Revised 15.11.2012 published: Print Citation Status MEDLINE |
| ISSN: | 1940-9818 |
| DOI: | 10.2144/0000113954 |