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US 12,270,028 B2Granted
Methods for seamless nucleic acid assembly
AssigneeTwist Bioscience Corporation
InventorsRebecca Nugent, Siyuan Chen, Elian Lee, Nathan Raynard
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Abstract
Provided herein are methods, systems, and compositions for seamless nucleic acid assembly. Methods, systems, and compositions as provided herein provide for efficient assembly of nucleic acids without primer removal. Methods, systems, and compositions for seamless nucleic acid assembly comprise use of an endonuclease or exonuclease, optionally in conjunction with additional enzymes to assemble nucleic acids or polynucleotides.
Claims20
A method for nucleic acid assembly, comprising:
- a. providing a plurality of polynucleotides, wherein each of the polynucleotides do not comprise a terminal region of sequence homology to another polynucleotide of the plurality of polynucleotides;
- and
- b. mixing the plurality of polynucleotides with a 3′ to 5′ exonuclease, a thermostable endonuclease, a high fidelity polymerase, and a thermostable ligase, wherein the plurality of polynucleotides are annealed in a processive predetermined order based on a complementary sequence between adjacent polynucleotides.
Dependencies are inferred from the claim text.