MODIFIED METHODS FOR DNA EXTRACTION FROM STRELITZIA REGINAE BANKS

由于鹤望兰组织内含有大量多糖类及多酚类物质 ,严重干扰DNA的抽提 ,利用常规的SDS法、CTAB法和高盐低pH值法都难以抽提出高质量的鹤望兰基因组DNA .本文报道了在进行若干预处理之后 ,再用常规的SDS、CTAB、SDS裂解的高盐低pH值法和CTAB裂解的高盐低pH值法提取基因组DNA的改进方法 .根据外观、琼脂糖凝胶电泳检测、D2 60nm/D2 80nm比值的测定、限制性酶切反应、PCR扩增的结果表明 ,用改进方法提取的基因组DNA无论在纯度上还是在完整性上都比常规方法要好 .其中改进SDS裂解的高盐低pH值法是提取鹤望兰基因组DNA的最好方法 .图 3表2参 18Strelitzia reginae Banks is rich in polysaccharides, polyphonels, and other secondary metabolites. Therefore, it is difficult to obtain high quality and quantity of genomic DNA from its tissues by conventional methods such as SDS method, CTAB method, and low pH medium with high salts method (LPHS). Modified procedures for DNA extraction from S. reginae are reported based on the above mentioned conventional methods, including the pretreatment of cell separation reagent to collect the nuclei and the use of β mercaptoethanol, polyvinylpyrrolidone (PVP) to inhibit the oxidization, and activated charcoal to clear the secondary metabolites, then followed by the original above mentioned methods to extract the total genomic DNA. The results showed that the modified methods were better than the original ones in terms of purity and totality of the genomic DNA. Among them, the LPHS by SDS cleavage is the best in quality, cost and simpleness of extraction. Fig 3, Tab 2, Ref 18厦门市园林局科技项目 (K990 2 )~