Mutagenesis method

The present invention provides a mutagenesis method wherein a nucleic acid molecule is mutagenized with at least one mutagenesis primer in a primer extension reaction and subsequently amplified by rolling circle amplification (RCA). The method involves the step of rendering the template strand unfav...

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Hauptverfasser: Brockmann Eeva-Christine, Lamminmäki Urpo, Huovinen Tuomas, Vehniäinen Markus
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creator Brockmann Eeva-Christine
Lamminmäki Urpo
Huovinen Tuomas
Vehniäinen Markus
description The present invention provides a mutagenesis method wherein a nucleic acid molecule is mutagenized with at least one mutagenesis primer in a primer extension reaction and subsequently amplified by rolling circle amplification (RCA). The method involves the step of rendering the template strand unfavorable for RCA. The method involves steps leading to selective amplification of only the mutated strand by a strand-displacing DNA polymerase. Multiple copies of the mutated plasmids are generated during multiple-primed RCA and the resulting DNA is transformed for use. The method is suitable for mutating both single-stranded and double-stranded DNA. The present invention also provides a kit for use in the mutagenesis method.
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subjects BEER
BIOCHEMISTRY
CHEMISTRY
COMPOSITIONS OR TEST PAPERS THEREFOR
COMPOSITIONS THEREOF
CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES
CULTURE MEDIA
ENZYMOLOGY
MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS
METALLURGY
MICROBIOLOGY
MICROORGANISMS OR ENZYMES
MUTATION OR GENETIC ENGINEERING
PROCESSES OF PREPARING SUCH COMPOSITIONS
PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS
SPIRITS
VINEGAR
WINE
title Mutagenesis method
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