Compositions and methods for enrichment of nucleic acids
First Claim
Patent Images
1. A method for enrichment of a target region in a DNA library comprising:
- a) providing a DNA library of double-stranded fragments with hairpin adapters on both ends, wherein the double-stranded fragments in the DNA library are not amplified nucleic acids, and wherein one or more of the double-stranded fragments are target fragments that comprise the target region;
b) subjecting the DNA library to endonuclease cleavage with an engineered endonuclease from a gene editing system that cleaves the target fragments at a first location to produce double-stranded ends, wherein the first location is present only once within at least one of the target fragments and is not within the target region;
c) linking stem-loop adapters to the double-stranded ends produced by the endonuclease cleavage, thereby forming asymmetric-adapter-ligated fragments, wherein the stem-loop adapters have a different sequence than the hairpin adapters; and
d) isolating the asymmetric-adapter-ligated fragments from other fragments in the reaction mixture that are not linked to the stem-loop adapters.
1 Assignment
0 Petitions
Accused Products
Abstract
Methods are provided for reducing the complexity of a population of nucleic acids prior to performing an analysis of the nucleic acids, e.g., sequence analysis. The methods result in a subset of the initial population enriched for a target region, which is typically located within one or more target fragments. The methods are particularly useful for analyzing populations having a high degree of complexity, e.g., chromosomal-derived DNA, whole genomic DNA, or mRNA populations.
-
Citations
27 Claims
-
1. A method for enrichment of a target region in a DNA library comprising:
-
a) providing a DNA library of double-stranded fragments with hairpin adapters on both ends, wherein the double-stranded fragments in the DNA library are not amplified nucleic acids, and wherein one or more of the double-stranded fragments are target fragments that comprise the target region; b) subjecting the DNA library to endonuclease cleavage with an engineered endonuclease from a gene editing system that cleaves the target fragments at a first location to produce double-stranded ends, wherein the first location is present only once within at least one of the target fragments and is not within the target region; c) linking stem-loop adapters to the double-stranded ends produced by the endonuclease cleavage, thereby forming asymmetric-adapter-ligated fragments, wherein the stem-loop adapters have a different sequence than the hairpin adapters; and d) isolating the asymmetric-adapter-ligated fragments from other fragments in the reaction mixture that are not linked to the stem-loop adapters. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27)
-
Specification