Multiple ligase compositions, systems, and methods
First Claim
1. A composition comprising a T4 DNA ligase mutant comprising the amino acid sequence of SEQ ID NO:
- 1.
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Abstract
Provided herein are compositions, systems, and methods using multiple ligases, wherein at least one of the ligases is an adenylation-deficient ATP-dependent ligase or an un-adenylated ATP-dependent ligase (e.g., present in an ATP free mixture). In certain embodiments, multiple ligases are used to ligate a pre-adenylated double stranded sequence to a non-adenylated double stranded sequence (e.g., the adenylation-deficient ATP-dependent ligase or un-adenylated ATP-dependent ligase ligates the first strand, and a second ligase ligates the second strand). In other embodiments, provided herein are mutant T4 ligases (e.g., K159S mutant or K159C mutant).
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5 Claims
- 1. A composition comprising a T4 DNA ligase mutant comprising the amino acid sequence of SEQ ID NO:
Specification