The present invention discloses a process for the preparation of variant polynucleotides using a combination of mutagenesis of a starting population of polynucleotides and recombination of the mutated polynucleotides. The process comprises the steps of subjecting a population of polynucleotides to (a series of) two (or more) separate PCR's comprising a first PCR with a forward mutation-specific primer for a position to be mutated and a reverse universal primer and a second PCR with a forward universal primer and a reverse mutation-specific primer for a position to be mutated. The products of the (series of) two (or more) PCR's are assembled by a polymerase, preferably in one tube.
申请公布号
WO03010183(A2)
申请公布日期
2003.02.06
申请号
WO2002EP08225
申请日期
2002.07.23
申请人
DSM N.V.;BOVENBERG, ROELOF, ARY, LANS;KERKMAN, RICHARD