Method for high-throughput AFLP-based polymorphism detection
DCFirst Claim
Patent Images
1. A method for determining dominant or co-dominant genotypes of one or more polymorphisms, comprising:
- (a) ligating an adaptor to a nucleic acid fragment to produce an adaptor-ligated nucleic acid fragment;
(b) amplifying the adaptor-ligated nucleic acid fragment to produce a library of amplified adaptor-ligated nucleic acid fragments, wherein the amplified adaptor-ligated nucleic acid fragments comprise an identifier tag;
(c) sequencing a pool of two or more libraries of amplified adaptor-ligated nucleic acid fragments using high-throughput sequencing, wherein each library comprises a unique identifier tag, wherein the sequencing comprises amplifying by PCR on a solid support;
(d) clustering the sequences per library and determining dominant or co-dominant genotypes of one or more polymorphisms in the two or more libraries using the identifier tag.
0 Assignments
Litigations
0 Petitions
Accused Products
Abstract
The invention relates to a method for the high throughput discovery, detection and genotyping of one or more genetic markers in one or more samples, comprising the steps of restriction endonuclease digest of DNA, adaptor-ligation, optional pre-amplification, selective amplification, pooling of the amplified products, sequencing the libraries with sufficient redundancy, clustering followed by identification of the genetic markers within the library and/or between libraries and determination of (co-)dominant genotypes of the genetic markers.
-
Citations
11 Claims
-
1. A method for determining dominant or co-dominant genotypes of one or more polymorphisms, comprising:
-
(a) ligating an adaptor to a nucleic acid fragment to produce an adaptor-ligated nucleic acid fragment; (b) amplifying the adaptor-ligated nucleic acid fragment to produce a library of amplified adaptor-ligated nucleic acid fragments, wherein the amplified adaptor-ligated nucleic acid fragments comprise an identifier tag; (c) sequencing a pool of two or more libraries of amplified adaptor-ligated nucleic acid fragments using high-throughput sequencing, wherein each library comprises a unique identifier tag, wherein the sequencing comprises amplifying by PCR on a solid support; (d) clustering the sequences per library and determining dominant or co-dominant genotypes of one or more polymorphisms in the two or more libraries using the identifier tag. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
-
-
11. A method for determining dominant or co-dominant genotypes of one or more polymorphisms, comprising:
-
(a) ligating an adaptor to a nucleic acid fragment to produce a library of adaptor-ligated nucleic acid fragment; (b) sequencing a pool of two or more libraries of adaptor-ligated nucleic acid fragments using high throughput sequencing, wherein each library comprises a unique identifier tag, and wherein the sequencing comprises amplifying by PCR on a solid support; (c) clustering the sequences per library and determining dominant or co-dominant genotypes of one or more polymorphisms in the two or more libraries using the identifier tag.
-
Specification