Directed evolution of DNA Polymerases : construction and screening of DNA Polymerase Mutant Libraries

Kurzanzeige
dc.contributor.authorGloeckner, Christiandeu
dc.contributor.authorKranaster, Ramon
dc.contributor.authorMarx, Andreas
dc.contributor.editorMahal, Lara
dc.contributor.editorRomesberg, Floyd
dc.contributor.editorShah, Kavita
dc.contributor.editorShamu, Caroline
dc.contributor.editorStrano, Michael S.
dc.contributor.editorThomas, Craig
dc.date.accessioned2011-06-16T08:43:13Zdeu
dc.date.available2011-06-16T08:43:13Zdeu
dc.date.issued2009
dc.description.abstractThe protocols in this article describe the construction of a mutant DNA polymerase library using error-prone PCR (epPCR) as a method for gene randomization, followed by screening of the library using two different approaches. The examples described use an N-terminally truncated form of the thermostable DNA polymerase I of Thermus aquaticus (Taq DNA polymerase), namely Klentaq (KTQ), and protocols are included for the identiÞcation of variants with (1) increased DNA lesion-bypass ability and (2) enhanced selectivity for DNA match/mismatch recognition. The screening assays are based on double-stranded DNA detection (using SYBR Green I) which can be carried out using standard laboratory equipment. The described assays are designed for use in a 384-well plate format to increase screening throughput and reduce material costs. For improved accuracy and ease of liquid handling, the use of an automated liquid handling device is recommended.eng
dc.description.versionpublished
dc.identifier.doi10.1002/9780470559277.ch090183deu
dc.identifier.urihttp://kops.uni-konstanz.de/handle/123456789/12600
dc.language.isoengdeu
dc.legacy.dateIssued2011-06-16deu
dc.rightsterms-of-usedeu
dc.rights.urihttps://rightsstatements.org/page/InC/1.0/deu
dc.subjectDNA polymerasedeu
dc.subjectdirected evolutiondeu
dc.subjectscreeningdeu
dc.subjectPCRdeu
dc.subjectprimer extensiondeu
dc.subject.ddc540deu
dc.titleDirected evolution of DNA Polymerases : construction and screening of DNA Polymerase Mutant Librarieseng
dc.typeJOURNAL_ARTICLEdeu
dspace.entity.typePublication
kops.citation.bibtex
@article{Gloeckner2009Direc-12600,
  year={2009},
  doi={10.1002/9780470559277.ch090183},
  title={Directed evolution of DNA Polymerases : construction and screening of DNA Polymerase Mutant Libraries},
  number={2},
  volume={2},
  journal={Current Protocols in Chemical Biology},
  pages={89--109},
  author={Gloeckner, Christian and Kranaster, Ramon and Marx, Andreas}
}
kops.citation.iso690GLOECKNER, Christian, Ramon KRANASTER, Andreas MARX, MAHAL, Lara, ed., Floyd ROMESBERG, ed., Kavita SHAH, ed., Caroline SHAMU, ed., Michael S. STRANO, ed., Craig THOMAS, ed., 2009. Directed evolution of DNA Polymerases : construction and screening of DNA Polymerase Mutant Libraries. In: Current Protocols in Chemical Biology. 2009, 2(2), pp. 89-109. Available under: doi: 10.1002/9780470559277.ch090183deu
kops.citation.iso690GLOECKNER, Christian, Ramon KRANASTER, Andreas MARX, MAHAL, Lara, ed., Floyd ROMESBERG, ed., Kavita SHAH, ed., Caroline SHAMU, ed., Michael S. STRANO, ed., Craig THOMAS, ed., 2009. Directed evolution of DNA Polymerases : construction and screening of DNA Polymerase Mutant Libraries. In: Current Protocols in Chemical Biology. 2009, 2(2), pp. 89-109. Available under: doi: 10.1002/9780470559277.ch090183eng
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kops.sourcefieldCurrent Protocols in Chemical Biology. 2009, <b>2</b>(2), pp. 89-109. Available under: doi: 10.1002/9780470559277.ch090183deu
kops.sourcefield.plainCurrent Protocols in Chemical Biology. 2009, 2(2), pp. 89-109. Available under: doi: 10.1002/9780470559277.ch090183deu
kops.sourcefield.plainCurrent Protocols in Chemical Biology. 2009, 2(2), pp. 89-109. Available under: doi: 10.1002/9780470559277.ch090183eng
kops.submitter.emailmichael.ketzer@uni-konstanz.dedeu
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source.periodicalTitleCurrent Protocols in Chemical Biology

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