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US 2014/0154676 A1Published application
High Fidelity Restriction Endonucleases
AssigneeNew England Biolabs, Inc.
InventorsZhenyu Zhu, Aine Quimby, Shengxi Guan, Dapeng Sun, Yishu Huang, Xuhui Lai, Siu-hong Chan, Xianghui Li, Shuang-Yong Xu, Chunhua Zhang
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Abstract
Methods and compositions are provided for engineering mutant enzymes with reduced star activity where the mutant enzymes have a fidelity index (FI) in a specified buffer that is greater than the FI of the non-mutated enzyme in the same buffer.
Claims5
1-74. (canceled)
A method of identifying a fidelity index (FI) of a restriction endonuclease and one or more variants thereof;
- comprising:
- selecting a reaction buffer and a DNA substrate containing the binding and cleavage site of the restriction endonuclease;
- permitting serially dilutions of the restriction endonuclease and the one or more variants thereof to cleave the DNA substrate;
- and
- determining an FI for each of the restriction endonuclease and the one or more variants thereof.
2 more elements
Dependencies are inferred from the claim text.