南方农业学报2012，Vol.43Issue(11)：1621-1625,5.DOI:10.3969/j:issn.2095-1191.2012.11.1621
适于SSR-PCR的农作物基因组总DNA高通量提取方法的建立
A method for high throughput DNA extraction from crop materials for SSR-PCR amplification
摘要
Abstract
[Objective]A novel,rapid,and universal method for DNA extraction from crops was developed to accelerate the studies on the genetic diversities and variations using various molecular markers.[Method] The DNA isolated from five crop varieties,viz.,sugarcane,rice,maize,peanut,and soybean using the method this paper account,and the total DNA was detected by agarose gel electrophoresis.Then PCR was performed using 2 pairs of SSR primer for each crop and the products were visualized on polyacrylamide gel electrophoresis.[Result]The presented method yielded pure DNA ranged from 100.0-160.0 g per gram of material depending upon the plant type.DNA samples with A260/A280 ratio ranged from 1.80 to 1.95.Furthermore,there was no visible RNA contamination in electrophoresed gels and the single bands of genomic DNA isolated from each crop were highly distinct.The DNA extracted from different crops showed high quality and reliability,therefore the DNA was ideal for marker applications.The PCR reactions using the simple sequence repeat (SSR) markers showed high repeatability,stability and the amplified bands were clear and distinct on the agarose gel.[Conclusion]For this DNA isolated method,the leaf pieces were put into PCR tubes followed by the addition of DNA extraction buffer,saved the process of grind using liquid nitrogen.Compared to other traditional DNA extraction methods and DNA isolation kits,this method is high throughput,simple,rapid,and very economic.关键词
甘蔗/水稻/玉米/花生/大豆/DNA提取/SSRKey words
sugarcane / rice / maize / peanut / soybean / DNA extraction / SSR分类
生物科学引用本文复制引用
梁俊,张妍,陈忠良,莫璋红,范业赓,吴凯朝,李鸣,李杨瑞..适于SSR-PCR的农作物基因组总DNA高通量提取方法的建立[J].南方农业学报,2012,43(11):1621-1625,5.基金项目
Key projects of Guangxi Natural Science Foundation (2011GXNSFD018021) （2011GXNSFD018021）
Postdoctoral Fund of Guangxi Academy of Agricultural Sciences (GUINONGKEBO2010017) （GUINONGKEBO2010017）
The Sci-tech Development Fund Programme of GXAAS(201002Z) （201002Z）
The Scientific Research and Technological Development Projects of Nanning(201102026B) （201102026B）
Natural Science Foundation of Guangxi (2010GXNSFD013034) （2010GXNSFD013034）
The Basic Scientific Research Fund of GXAAS (Guinongke2012YM11) （Guinongke2012YM11）
