从顽拗植物荔枝中提取基因组DNA技术的研究

STUDY ON GENOMIC DNA EXTRACTION FROM RECALCITRANT LITCHI

针对荔枝体内含有大量影响DNA提取质量的多酚等次生代谢物质的特点 ,在常规SDS和CTAB提取方法的基础上做了技术改进 ,即在核裂解之前先破碎细胞 ,将存在于细胞质中的次生物质除去后再裂解细胞核 ,同时加入活性炭以吸附杂质 ,反复清洗几次后加入核裂解液释放基因组DNA ,纯化之后经外观和电泳检测 ,以及D2 60nm和D2 80nm比值的测定 .限制性内切酶反应 ,PCR扩增的结果表明 ,用改进方法提取的DNA无论在纯度上还是在完整性上都比用常规方法要好 ,用改进的CTAB法虽然所得的DNA在产量上要少 ,但质量比其它方法更好 .图 2表 1参

Based on the influence of polyphenols in litchi on extraction quantity of genomic DNA, an efficient procedure was developed for DNA extraction from litchi by modifying the original SDS and CTAB methods. The major improvement included collection of nuclei before extracting DNA and adding activated charcoal to bind the secondary compounds which were purified with DNA. After nuclei washed for several times, extraction solution was added to extract genomic DNA. The results from electrophoresis, ratios of D 260nm and D 280nm , digestion of restriction endonucleases and PCR amplification suggested that the modified methods, especially the modified CTAB method produce high quality DNA compared with conventional methods. Fig 2, Tab 1, Ref 7