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目的:本研究采用改良CTAB法提取重楼属3种植物的DNA,分析了CTAB浓度对其DNA得率以及醋酸钾(KAc)或KAc+20%乙醇对其DNA纯度的影响,为重楼属植物药材的分子标记鉴定研究提供技术和理论依据。方法:采用基于CTAB的6种方法提取重楼属3种植物干燥根茎的DNA,并用紫外分光光度法测定A260和A280的值计算评价DNA得率和纯度。结果:4%CTAB提取3种重楼的DNA得率均高于2%CTAB;添加KAc的方法 2(2%CTAB)、方法 5(4%CTAB)以及添加KAc+20%乙醇的方法 3(2%CTAB)和方法 6(4%CTAB)提取的DNA纯度和质量以及通用引物ITS2F/ITS2R的PCR扩增结果均优于不添加KAc或KAc+20%乙醇的方法 1和方法 4。结论:本研究表明4%CTAB有利于重楼属3种植物干燥根茎DNA得率的提高,在DNA沉淀之前添加KAc或KAc+20%乙醇有利于改善DNA纯度和质量。
Objective: In this study, the improved CTAB method was used to extract the DNA of three species of Rhizoma Paridis. The effects of CTAB concentration on DNA yield and DNA purity of KAc or KAc + 20% ethanol were analyzed. Plant molecular identification of medicinal herbs to provide technical and theoretical basis. Methods: Six kinds of methods based on CTAB were used to extract DNA from the rhizomes of three species of Rheum. The DNA yield and purity of DNA were calculated by UV spectrophotometry. Results: The yields of DNA extracted from 3 kinds of reed houses by 4% CTAB were higher than that of 2% CTAB. Method 2 (2% CTAB) with KAc added, method 5 (4% CTAB) and KAc + 20% 2% CTAB) and method 6 (4% CTAB), and PCR amplification results of the universal primer ITS2F / ITS2R were superior to those of methods 1 and 4 without addition of KAc or KAc + 20% ethanol. Conclusion: This study shows that 4% CTAB is conducive to the increase of DNA yield of dried rhizomes of 3 species of Rhodopes, and the addition of KAc or KAc + 20% ethanol before DNA precipitation is helpful to improve DNA purity and quality.