Method for directed evolution by random chimeragenesis on transient templates

摘要

A method is provided wherein at least one template is used to align oligonucleotides of interest or fragments thereof to generate chimeric polynucleotides. The oligonucleotides are allowed to hybridize to the template or templates. If necessary, unhybridized termini of hybridized oligonucleotides are removed and gaps between hybridized oligonucleotides are filled. Adjacently hybridized oligonucleotides are ligated to generate chimeric polynucleotides hybridized to the template or templates. The chimeric polynucleotides can then be subjected to selection and/or screening for desired characteristics. Prior to or concomitant with selection/screening, the template is optionally removed.