药用野生稻基因组DNA提取方法比较

Comparison of Different Extraction Methods of Genomic DNA from Oryza officinalis

以药用野生稻为试验材料,分别选用传统的CTAB提取法、CTAB改良方法、碱裂解法、磁珠法、吸附柱法、尿素提取法及酶消化法7种方法提取水稻叶片组织DNA,7种方法分别标记为A~G。通过比较这些方法提取出DNA的浓度和纯度,分析这些方法的优劣势。7种方法均能获得较好的基因组,按获得DNA浓度表现为A>B>F>C>G>E>D,纯度表现为D>E>C>G>B>F>A。综合考量DNA的纯度、浓度、提取时间、成本、安全无毒等方面因素,若对基因组的质量要求不高,只是做简单的检测且考虑成本问题,可以选择CTAB提取法、CTAB改良方法和尿素提取法;若需要高纯度基因组,可以选择磁珠法、吸附柱法、碱裂解法;若考虑安全无毒、快速提取及成本可控,可以选择磁珠法、吸附柱法;若样品稀有,可以选用CTAB提取法和CTAB改良法。

With Oryza officinalis as test material,seven methods were used to extract the genomic DNA from the Oryza officinalis,including traditional CTAB method,improved CTAB method,alkaline lysis method,magnetic bead method,adsorption column method,urea extraction method and enzyme digestion method.The seven methods are labeled A-G,respectively.By comparing the concentration and purity of DNA extracted by these methods,the advantages and disadvantages of these methods were analyzed.All seven methods can obtain genomic DNA.The order of concentration from high to low is A>B>F>C>G>E>D,and the order of purity from high to low is D>E>C>G>B>F>A.Considering the purity,concentration,extraction time,cost,safety and non⁃toxicity of the extraction process,CTAB or CTAB improvement extraction and urea extraction method may be preferred according to the cost and purity.The magnetic bead,adsorption column and alkaline lysis method is preferred when high⁃purity are required.The magnetic bead and adsorption column method is preferred when safety,non⁃toxicity,rapid extrac⁃tion and cost little are required,while the CTAB and CTAB modification method is recommended for limited samples.