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báo cáo khoa học: " Simultaneous mutation detection of three homoeologous genes in wheat by High Resolution Melting analysis and Mutation Surveyor®"

Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: Simultaneous mutation detection of three homoeologous genes in wheat by High Resolution Melting analysis and Mutation Surveyor® | BMC Plant Biology BioMed Central Open Access Methodology article Simultaneous mutation detection of three homoeologous genes in wheat by High Resolution Melting analysis and Mutation Surveyor Chongmei Dong 1 Kate Vincent1 2 and Peter Sharp1 Address 1Plant Breeding Institute University of Sydney PMB 4011 Narellan NSW 2567 Australia and 2Australian Centre for Plant Functional Genomics PMB 1 Glen Osmond SA 5064 Australia Email Chongmei Dong - chongmei.dong@sydney.edu.au KateVincent-kate.vincent@sydney.edu.au Peter Sharp - peter.sharp@sydney.edu.au Corresponding author Published 4 December 2009 Received 21 May 2009 BMC Plant Biology 2009 9 143 doi l0.ll86 l47l-2229-9-l43 Accepted 4 December 2009 This article is available from http www.biomedcentral.cOm l47l-2229 9 l43 2009 Dong et al licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License http creativecommons.org licenses by 2.0 which permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited. Abstract Background TILLING Targeting Induced Local Lesions IN Genomes is a powerful tool for reverse genetics combining traditional chemical mutagenesis with high-throughput PCR-based mutation detection to discover induced mutations that alter protein function. The most popular mutation detection method for TILLING is a mismatch cleavage assay using the endonuclease CelI. For this method locus-specific PCR is essential. Most wheat genes are present as three similar sequences with high homology in exons and low homology in introns. Locus-specific primers can usually be designed in introns. However it is sometimes difficult to design locus-specific PCR primers in a conserved region with high homology among the three homoeologous genes or in a gene lacking introns or if information on introns is not available. Here we describe a mutation detection method which combines High Resolution Melting HRM .