华北农学报2017，Vol.32Issue(6)：67-72,6.DOI:10.7668/hbnxb.2017.06.010
大白菜基因组DNA快速提取方法的研究
Study on Rapid Extraction of Genomic DNA from Chinese Cabbage
摘要
Abstract
Rapid and efficient DNA extraction is the key step in large-scale molecular breeding of crop. To con-struct a method for rapid extraction of genomic DNA from Chinese cabbage, the Chinese cabbage leaves were used as experimental materials, the quality of DNA extracted by the CTAB method, two-step CTAB method and four alkaline lysis methods was compared, the PCR amplification effects of DNA extracted by different methods were analyzed, and the preservation time and preservation conditions of genomic DNA extracted by different methods were compared, the optimal method was selected for the application of molecular marker-assisted selection against clubroot disease. The results showed that genomic DNA extracted by the CTAB and three kinds of alkaline lysis methods could be used as a template for PCR, and the PCR amplification products could be detected by 8％ non-denaturing polyacrylamide gel e-lectrophoresis to clear bands. Among them, the alkaline lysis methodⅢ, not only the extraction quality was good and the extraction process was simple and fast, but also it could meet the needs of high-throughput DNA extraction of Chi-nese cabbage, the extracted DNA was stored at 4℃ and -20℃, and the genomic DNA stored for 30 days was used as a template for PCR amplification, the products was still detected by polyacrylamide gel electrophoresis to clear bands, it illustrates that this method has a long preservation time and it has been proved that this method has a good effect on the application of molecular marker-assisted selection against clubroot disease. Alkaline lysis methodⅢhas significantly improved the efficiency of Chinese cabbage molecular marker-assisted selection, and can be widely used in Chinese cabbage molecular marker-assisted selection breeding.关键词
大白菜/基因组DNA/快速提取Key words
Chinese cabbage/Genomic DNA/Rapid extraction分类
农业科技引用本文复制引用
王涛,王超楠,张红,温娟娟,张斌..大白菜基因组DNA快速提取方法的研究[J].华北农学报,2017,32(6):67-72,6.基金项目
国家重点研发计划项目( 2017YFD0101801 ) （ 2017YFD0101801 ）
天津市自然科学基金项目( 16JCYBJC29300 ) （ 16JCYBJC29300 ）
国家重点研发计划项目(2016YFD0101701) （2016YFD0101701）
天津市现代农业产业技术体系创新团队建设专项计划(ITTVRS2017003) （ITTVRS2017003）
国家大宗蔬菜产业技术体系(CARS-25-G-02) （CARS-25-G-02）
天津市科技计划项目(16PTSYJC00260) （16PTSYJC00260）
