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

第一作者： Chu, Zhi-Zhan

作者： Chu, Zhi-Zhan;Liu, Yao-Guang;Li, Yi-Dan;Liu, Xiang-Guo;Hao, Dong-Yun;Jing, Hai-Chun

作者机构：

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.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序