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US 12,644,115 B2Granted
Methylation-mediated adapter removal on nucleic acid sequences
AssigneeTwist Bioscience Corporation
InventorsSiyuan Chen, Xuan Yu Elian Lee, David Lin, Daniel J. Ramirez, Tavneet Gill, Patrick Cherry, Esteban Toro
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Abstract
Provided herein are methods, systems, and compositions for efficient nucleic acid assembly with improved representation and distribution. Provided herein are methods, systems, and compositions for efficient nucleic acid assembly of nucleic acids encoding genes for use in various downstream processes.
Claims20
A method for nucleic acid assembly, comprising:
- providing a plurality of polynucleotides, wherein each of the polynucleotides comprises:
- (i) a 5′ adapter sequence comprising at least one Type IIS endonuclease site;
- or
- (ii) a 3′ adapter sequence comprising at least one Type IIS endonuclease site;
- amplifying the plurality of polynucleotides using a reaction mixture comprising about 5% to about 100% modified bases;
- and
- mixing the plurality of polynucleotides with a Type IIS restriction enzyme to generate a plurality of nucleic acids.
4 more elements
A method for adapter removal, comprising:
- providing a plurality of polynucleotides encoding a gene comprising one or more modifications associated with a disease or disorder, wherein each of the polynucleotides comprises:
- (i) a 5′ adapter sequence comprising at least one Type IIS endonuclease site;
- or
- (ii) a 3′ adapter sequence comprising at least one Type IIS endonuclease site;
- amplifying the plurality of polynucleotides using a reaction mixture comprising about 5% to about 100% modified bases;
- and
- mixing the plurality of polynucleotides with a Type IIS restriction enzyme to generate a plurality of nucleic acids.
4 more elements
A method for adapter removal, comprising:
- providing a plurality of polynucleotides encoding a gene comprising one or more modifications associated with a disease or disorder, wherein each of the polynucleotides comprises:
- (i) a 5′ adapter sequence comprising at least one Type IIS endonuclease site;
- and
- (ii) a 3′ adapter sequence comprising at least one Type IIS endonuclease site;
- amplifying the plurality of polynucleotides using a reaction mixture comprising about 5% to about 100% modified bases;
- and
- mixing the plurality of polynucleotides with a Type IIS restriction enzyme to generate a plurality of nucleic acids.
4 more elements
Dependencies are inferred from the claim text.