METHOD FOR HIGH-THROUGHPUT AFLP-BASED POLYMORPHISM DETECTION

摘要

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.

主权项

1. A method for genotyping one or more genetic markers, comprising:
(a) ligating at least one adaptor to at least one restriction fragment to produce at least one adaptor-ligated restriction fragment; (b) amplifying by solution PCR the at least one adaptor-ligated restriction fragment with at least one primer pair to produce a library of at least one amplified adaptor-ligated double stranded restriction fragment, wherein the amplified adaptor-ligated restriction fragment comprises an identifier tag; (c) sequencing a pool of two or more libraries of tagged amplified adaptor-ligated restriction fragments using high throughput sequencing technology, wherein each library comprises a unique identifier tag, wherein the sequencing comprises amplifying by PCR on a solid support; (d) clustering the sequences; and (e) identifying genetic markers within the library and/or between libraries using the identifier tag.