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

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

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GLOECKNER, 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. 2(2), pp. 89-109

@article{Gloeckner2009Direc-12600, title={Directed evolution of DNA Polymerases : construction and screening of DNA Polymerase Mutant Libraries}, year={2009}, doi={10.1002/9780470559277.ch090183}, number={2}, volume={2}, journal={Current Protocols in Chemical Biology}, pages={89--109}, author={Gloeckner, Christian and Kranaster, Ramon and Marx, Andreas} }

<rdf:RDF xmlns:rdf="http://www.w3.org/1999/02/22-rdf-syntax-ns#" xmlns:bibo="http://purl.org/ontology/bibo/" xmlns:dc="http://purl.org/dc/elements/1.1/" xmlns:dcterms="http://purl.org/dc/terms/" xmlns:xsd="http://www.w3.org/2001/XMLSchema#" > <rdf:Description rdf:about="https://kops.uni-konstanz.de/rdf/resource/123456789/12600"> <dc:creator>Marx, Andreas</dc:creator> <dc:contributor>Strano, Michael S.</dc:contributor> <dc:contributor>Thomas, Craig</dc:contributor> <dcterms:rights rdf:resource="http://nbn-resolving.org/urn:nbn:de:bsz:352-20140905103605204-4002607-1"/> <dc:contributor>Mahal, Lara</dc:contributor> <dc:contributor>Kranaster, Ramon</dc:contributor> <dcterms:issued>2009</dcterms:issued> <dc:contributor>Shamu, Caroline</dc:contributor> <bibo:uri rdf:resource="http://kops.uni-konstanz.de/handle/123456789/12600"/> <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-06-16T08:43:13Z</dcterms:available> <dcterms:abstract xml:lang="eng">The 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.</dcterms:abstract> <dc:contributor>Gloeckner, Christian</dc:contributor> <dc:rights>deposit-license</dc:rights> <dcterms:title>Directed evolution of DNA Polymerases : construction and screening of DNA Polymerase Mutant Libraries</dcterms:title> <dc:contributor>Romesberg, Floyd</dc:contributor> <dc:creator>Kranaster, Ramon</dc:creator> <dc:contributor>Shah, Kavita</dc:contributor> <dc:language>eng</dc:language> <dc:creator>Gloeckner, Christian</dc:creator> <dc:contributor>Marx, Andreas</dc:contributor> <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-06-16T08:43:13Z</dc:date> </rdf:Description> </rdf:RDF>

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