A Cost-effective High-resolution Melting Approach using the EvaGreen Dye for DNA Polymorphism Detection and Genotyping in Plants
文献类型： 外文期刊
作者： Li, Yi-Dan 2 ; Chu, Zhi-Zhan 1 ; Liu, Xiang-Guo 2 ; Jing, Hai-Chun 3 ; Liu, Yao-Guang 1 ; Hao, Dong-Yun 2 ;
作者机构： 1.S China Agr Univ, Coll Life Sci, Guangzhou 510642, Guangdong, Peoples R China
2.Jilin Acad Agr Sci, Biotechnol Res Ctr, Changchun 130033, Peoples R China
3.Chinese Acad Sci, Inst Bot, Beijing 100093, Peoples R China
期刊名称：JOURNAL OF INTEGRATIVE PLANT BIOLOGY （ 影响因子：7.061； 五年影响因子：6.002 ）
ISSN： 1672-9072
年卷期： 2010 年 52 卷 12 期
页码：
收录情况： SCI
摘要： 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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