Compositions and methods of targeted nucleic acid enrichment by loop adapter protection and exonuclease digestion
AssigneeKing Abdullah University of Science and Technology
InventorsMo Li, Chongwei Bi, Lin Wang
Life of this patent
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Abstract
Compositions and methods for enriching DNA from any loci of interest are provided. Endonuclease(s) such as RNA-guided nucleases Cpf1/Cas9 can cut the flanking regions of the gene of interest in, for example genomic DNA, followed by ligation of loop adapters. The loop adapters protect the gene of interest for subsequent digestion with one or more exonucleases. Multiple genes of interest can be digested and ligated with loop adapters at the same time. The undigested target sequence can be further purified by, for example, gel extraction using, for example, a commercial DNA purification kit. The enriched DNA can be used for sequencing.
Claims18
A nucleic acid loop adapter comprising
- (i) a sticky end nucleic acid segment designed to hybridize with a sticky end of a target linear nucleic acid;
- (ii) a first hybridization nucleic acid segment designed to hybridize with a second hybridization nucleic acid segment;
- (iii) a loop nucleic acid segment that remains single stranded;
- and
- (iv) the second hybridization nucleic acid segment;
- wherein the first hybridization nucleic acid segment is sequence CAGTCCAGG (SEQ ID NO:
- 2), and the second hybridization nucleic acid segment is sequence CCTGGACTG (SEQ ID NO:
- 3);
- or wherein the second hybridization nucleic acid segment is sequence CAGTCCAGG (SEQ ID NO:2), and the first hybridization nucleic acid segment is sequence CCTGGACTG (SEQ ID NO:3);
- or
- wherein the second hybridization nucleic acid segment consists of the sequence CAGTCCAGG (SEQ ID NO:2), and the first hybridization nucleic acid segment consists of the sequence CCTGGACTG (SEQ ID NO:3).
A nucleic acid loop adapter having the nucleic acid sequence of SEQ ID NO:
- 1.
Dependencies are inferred from the claim text.