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DHPLC Technology as a High-throughput Detection of Mutations in a Durum Wheat TILLING Population
EP22696
Poster Title: DHPLC Technology as a High-throughput Detection of Mutations in a Durum Wheat TILLING Population
Submitted on 02 Feb 2015
Author(s): Colasuonno P.1, Incerti O. 1, Lozito M.L. 1, Sbalzarini M. 2, Zaccagna P. 2, Papadimitriou S. 2, Blanco A. 1, Gadaleta A. 1
Affiliations: 1Department of Soil, Plant and Food Sciences, sect. Genetics and Plant Breeding, University of Bari, Via Amendola 165/A, 70126 Bari (Italy) 2 Transgenomics Omaha, NE, USA
This poster was presented at PAG 2015
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Poster Information
Abstract: Durum wheat (Triticum turgidum ssp. durum) has a tetraploid genome (AABB genome) with genes in two homoeologous copies, a high proportion of repetitive DNA regions and a low gene density. Current tools for the discovery of sequence alterations in target genes rely on reverse genetic approaches such as the TILLING (Targeting Induced Local Lesions IN Genomes) technique. TILLING is based on the detection of chemical induced mutations in heteroduplex mainly using endonuclease (CelI enzyme), standard gel electrophoresis, fluorescent capillary electrophoresis and/or LI-COR techniques. Alternatively, DHPLC approach reveals the presence of genetic variation by the differential retention of the homo- and heteroduplex DNA molecules on ion-pair reversed-phase high-pressure liquid chromatography supported under partial denaturing conditions.Summary: This study is a beautiful example of DHPLC technology application and shows an alternative tool to current strategies of SNP detection based on genotyping array.Report abuse »
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