不同种质荔枝基因组DNA提取研究

Extraction of genomic DNA in different litchi germplasm

针对荔枝体细胞富含多酚、多糖、色素以及蛋白质等次生物质的特点,以荔枝叶片为材料,分别采用CTAB法和SDS法对荔枝野生、半野生和栽培种质的基因组DNA进行提取。结果表明,SDS法提取的荔枝基因组DNA效果稳定,获得DNA的机率为100%,DNA纯度高、质量好,可用于AFLP酶切、连接、预扩增和选择性扩增,并获得清晰、稳定的扩增产物;CTAB法提取的荔枝基因组DNA效果不稳定,DNA浓度较低,易出现多糖等次生物质与DNA共沉淀现象,无法检测出DNA。因此,SDS法可用于不同荔枝种质基因组DNA的提取,进行AFLP、SSR、ISSR分析。

In accordance with large amounts of secondary metabolites such as polyphenol, polysaccharide, pigment and protein, and so on in leaf cells of litchi, the genomic DNA of wild, semi-wild and cultivated verieties of litchi were extracted by SDS and CTAB methods. The results showed that the effects of SDS extraction on genomic DNA of litehi were stable, the genomic DNA with high-quality and high-purity could be obtained and used for AFLP analysis, and the amplified products were clear and stable. The effects of CTAB extraction on genomic DNA were unstable, and DNA could not be detected at Dower concentration and DNA coprecipitated with secondary metabolites. Therefore, SDS method could be used for extracting genomic DNA of different litchi germplasm for AFLP, SSR and ISSR analysis.