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 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.
申请公布号
EP2516643(A1)
申请公布日期
2012.10.31
申请号
EP20100838753
申请日期
2010.12.21
申请人
TURUN YLIOPISTO
发明人
HUOVINEN, TUOMAS;LAMMINMAEKI, URPO;BROCKMANN, EEVA-CHRISTINE;VEHNIAEINEN, MARKUS