茶树基因组DNA的高效提取方法

Efficient Methods for Genomic DNA Extraction from Tea Plant

为了从富含多酚和多糖等多种次生代谢物质的茶树叶片中分离出高质量的基因组 DNA,对 4种植物 DNA提取方法进行改良后 ,以茶树春梢为材料 ,对提取效果进行了比较 ,并首次采用 CTAB区室法提取茶树基因组DNA.结果表明 ,用该 4种方法提取的 DNA,其 OD2 6 0 / OD2 80 都在 1.8以上 ,相对分子质量均大于 2 1kb,能完全被Eco R 酶切消化 ,也能获得清晰的 PCR扩增图谱 .因此 ,只要操作合理 ,4种方法均能提取出高质量的 DNA.

In order to isolate high quality genomic DNA from tea plant leaves enriching polyphenols and polysaccharides, this paper is an attempt to compare the DNA extracting effect of four modified methods including CTAB general method, CTAB subarea method,SDS general method and SDS subarea method.Besides the CTAB subarea method is used for the first time for genomic DNA isolation from tea plant.Results show that the value of OD 260 / OD 280 for all the DNA samples obtained with these four modified methods is over 1.8, the molecular weight of DNAs is more than 21 kb, and DNAs are completely digestible with restriction endonucleases and amplifiable in the polymerase chain reaction (PCR), indicating freedom from common contaminating compounds. The high quality genomic DNA can be obtained with any method if the process is reasonable.