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US 12,529,100 B2Granted
Methods for removal of adaptor dimers from nucleic acid sequencing preparations
AssigneeRoche Sequencing Solutions, Inc.
InventorsArkadiusz Bibillo, Randall W. Davis
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Abstract
Sequencing adaptors and methods are provided for preparation of polynucleotides for sequencing. The sequencing adaptors contain a portion of a recognition sequence for a methyl-dependent endonuclease. Unwanted adaptor dimers that form during ligation of adaptors to target polynucleotides produce a complete restriction sequence and are cleaved by the endonuclease, followed by exonuclease digestion, thereby removing the dimers.
Claims3
An adaptor for polynucleotide sequencing, wherein the adaptor comprises:
- a double-stranded polynucleotide duplex,
- wherein one end of the double-stranded polynucleotide duplex comprises a part of a complete recognition sequence for a methyl-dependent endonuclease,
- wherein a complete recognition sequence for a methyl-dependent endonuclease will be formed if two adaptors are covalently linked together to produce an adaptor dimer, such that the part of the complete recognition sequence for the methyl-dependent endonuclease of the first adaptor is adjacent to the part of the complete recognition sequence of the second adaptor, to generate the complete recognition sequence for a methyl-dependent endonuclease, wherein the complete recognition sequence consists of a cleavage site for a methyl-dependent endonuclease,
- wherein the cleavage site comprises:
- (i) a nucleic acid sequence of CCMeGG that is recognized by the methyl-dependent endonuclease MspI or MspII, and/or (ii) a nucleic acid sequence of GAMeTC that is recognized by the methyl-dependent endonuclease DpnI or DpnII, and
- wherein the methyl-dependent endonuclease is selected from a group consisting of:
- MspI, MspII, DpnI, and DpnII.
4 more elements
Dependencies are inferred from the claim text.