Methods for Generating Pools of Variants of a DNA Template

摘要

The invention provides methods for generating pools of variants of DNA templates, and methods of using pools of variants to identify sequences involved in conferring sensitivity or resistance to environmental factors.

主权项

1. A method of generating a mixture of variants of a template DNA molecule, said method comprising:
a) providing a plurality of oligonucleotides, wherein each of said oligonucleotides:
i) corresponds to a first region of said template DNA molecule, andii) has at least one sequence variation as compared to the sequence of said first region of said template DNA molecule; b) providing a plurality of linearized plasmid vectors comprising the remainder of said template DNA molecule, but lacking said first region of said template DNA molecule at the ends of said linearized plasmid vectors; and c) joining said plurality of oligonucleotides to said plurality of linearized plasmid vectors, thereby generating a mixture of plasmids comprising a mixture of variants of said first region of said template DNA molecule.