DNA polymerase profiling

Kurzanzeige
dc.contributor.authorSummerer, Daniel
dc.date.accessioned2011-06-29T12:41:22Zdeu
dc.date.available2011-06-29T12:41:22Zdeu
dc.date.issued2008
dc.description.abstractWe report a simple homogeneous fluorescence assay for quantification of DNA polymerase function in high throughput. The fluorescence signal is generated by the DNA polymerase triggering opening of a molecular beacon extension of the template strand. A resulting distance alteration is reported by fluorescence resonance energy transfer between two dyes introduced into the molecular beacon stem. We describe real-time reaction profiling of two model DNA polymerases. We demonstrate kinetic characterization, rapid optimization of reaction conditions, and inhibitor profiling using the presented assay. Furthermore, to supersede purification steps in screening procedures of DNA polymerase mutant libraries, detection of enzymatic activity in bacterial expression lysates is described.eng
dc.description.versionpublished
dc.format.mimetypeapplication/pdfdeu
dc.identifier.citationFirst publ. in: Methods in molecular biology 429 (2008), pp. 225-235deu
dc.identifier.doi10.1007/978-1-60327-040-3_16deu
dc.identifier.pmid18695970
dc.identifier.ppn34672922Xdeu
dc.identifier.urihttp://kops.uni-konstanz.de/handle/123456789/353
dc.language.isoengdeu
dc.legacy.dateIssued2011deu
dc.relation.ispartofseriesMethods in Molecular Biology
dc.rightsterms-of-usedeu
dc.rights.urihttps://rightsstatements.org/page/InC/1.0/deu
dc.subject.ddc540deu
dc.subject.gndHigh throughput screeningdeu
dc.subject.gndFRETdeu
dc.subject.gndDNA polymerasedeu
dc.subject.gnddirected evolutiondeu
dc.subject.gndinhibitor screeningdeu
dc.subject.gndfluorescent probesdeu
dc.titleDNA polymerase profilingeng
dc.typeJOURNAL_ARTICLEdeu
dspace.entity.typePublication
kops.citation.bibtex
@article{Summerer2008polym-353,
  year={2008},
  doi={10.1007/978-1-60327-040-3_16},
  title={DNA polymerase profiling},
  volume={429},
  issn={1064-3745},
  journal={Methods in molecular biology (Clifton, N.J.)},
  pages={225--235},
  author={Summerer, Daniel}
}
kops.citation.iso690SUMMERER, Daniel, 2008. DNA polymerase profiling. In: Methods in molecular biology (Clifton, N.J.). 2008, 429, pp. 225-235. ISSN 1064-3745. Available under: doi: 10.1007/978-1-60327-040-3_16deu
kops.citation.iso690SUMMERER, Daniel, 2008. DNA polymerase profiling. In: Methods in molecular biology (Clifton, N.J.). 2008, 429, pp. 225-235. ISSN 1064-3745. Available under: doi: 10.1007/978-1-60327-040-3_16eng
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kops.sourcefieldMethods in molecular biology (Clifton, N.J.). 2008, <b>429</b>, pp. 225-235. ISSN 1064-3745. Available under: doi: 10.1007/978-1-60327-040-3_16deu
kops.sourcefield.plainMethods in molecular biology (Clifton, N.J.). 2008, 429, pp. 225-235. ISSN 1064-3745. Available under: doi: 10.1007/978-1-60327-040-3_16deu
kops.sourcefield.plainMethods in molecular biology (Clifton, N.J.). 2008, 429, pp. 225-235. ISSN 1064-3745. Available under: doi: 10.1007/978-1-60327-040-3_16eng
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