| A cost-effective high-resolution melting approach using the EvaGreen dye for DNA polymorphism detection and genotyping in plants. | |
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MedLine Citation:
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PMID: 21106003 Owner: NLM Status: In-Process |
Abstract/OtherAbstract:
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High-resolution melting (HRM) analysis relies on the use of fluorescent dyes, such as LCGreen, ResoLight, and SYTO9, which bind in a saturated manner to the double-stranded DNAs. These dyes are expensive in use and may not be affordable when dealing with a large quantity of samples. EvaGreen is a much cheaper DNA helix intercalating dye and has been used in quantitative real-time polymerase chain reaction (PCR) and post-PCR DNA melt curve analysis. Here we report on the development of an EvaGreen-based HRM analysis and its performance, in comparison with the popular LCGreen-based HRM analysis, in detection of DNA polymorphism in plants. We found that various polymorphisms ranged from single nucleotide polymorphisms (SNPs) to Indels were equally detected by using EvaGreen- or LCGreen-based HRM. EvaGreen dye was sensitive enough in discovery of SNPs in fivefold pooled samples. Using this economical dye we successfully identified multiple novel mutant alleles of Gln1-3 gene, which produces a cytosolic glutamine synthetase isoenzyme (GS1), in a maize ethyl methanesulfonate (EMS)-mutagenized library, and genotyped rice mapping populations with SNP markers. The current results suggest that EvaGreen is a promising dye for HRM analysis for its ease to use and cost effectiveness. |
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Authors:
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Yi-Dan Li; Zhi-Zhan Chu; Xiang-Guo Liu; Hai-Chun Jing; Yao-Guang Liu; Dong-Yun Hao |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Journal of integrative plant biology Volume: 52 ISSN: 1744-7909 ISO Abbreviation: J Integr Plant Biol Publication Date: 2010 Dec |
Date Detail:
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Created Date: 2010-11-25 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101250502 Medline TA: J Integr Plant Biol Country: China (Republic : 1949- ) |
Other Details:
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Languages: eng Pagination: 1036-42 Citation Subset: IM |
Copyright Information:
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© 2010 Institute of Botany, Chinese Academy of Sciences. |
Affiliation:
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Biotechnology Research Centre, Jilin Academy of Agricultural Sciences, Changchun 130033, China. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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