三种提取冬虫夏草菌丝体基因组DNA方法的比较.PDF

生物技术通报 BIO TE CHN OLOG Y B ULLETIN 20 10 8 DNA 谢放 陈京津 朱子雄 ( , 730070) : 采用改良氯化苄法CTAB法蛋白酶 K裂解法这三种方法, 对冬虫夏草菌 体基因组 DNA 进行提取, 将提取 的基因组 DNA 经过紫外分光光度计检测纯度和计算浓度琼脂糖凝胶电泳分析及 PCR扩增, 结果表明, 3种方法均能提取到 符合分子生物学的 DNA, 其中蛋白酶 K裂解法纯度最高, 含蛋白质少, 改良氯化苄法CTAB 法次之而浓度则是改良氯化苄 法最高, CTAB 法蛋白酶 K 裂解法次之3种方法所提取的 DNA 均能扩增出理想条带 : 冬虫夏草 菌 体 基因组 DNA 提取方法 Comparison of ThreeM ethods for Extracting Genom ic DNA ofCordy cep s sinensis M ycelium X ie Fang Chen Jingjin Zhu Z i iong (College of Chem icaland BiologicalEngineering, Lanzhou JiaoTong University,Lanzhou 730070) Abstrac:t Threem ethods ofm odified benzyl ch loride method, CTAB m ethod and proteinase K lysis m ethod w ere used to e tract the DNA of cordyceps m ycelium, the concen tration of DNA w as detected by UV spectrophotom eter and calculated the DNA purity, agar ose condensate gel electrophoresis and PCR amp lification w as analyzed. The resu lts showed that, the threem ethods w ere able to e tract the DNA from Cordyceps m ycelium, the DNA w hich w as e tracted by proteinase K lysis m ethod was the h ighest purity, contain ing less protein and them od ified benzyl chloridem ethod and the CTAB m ethod followed. The DNA wh ich w as e tracted by the benzyl chloride m ethod w as the highest concentration, the CTAB method follow ed, then the proteinaseK lysis method. The DNA w hich was e tracted by three kinds ofm ethod could amp lify the ideal bands. K ey w ords: Cordyceps sinensis M ycelium Genom ic DNA E traction m ethod DNA , , , PCR , , 16 45 d 12 主要试剂 , , (, , , ); SDS(); EDTA (, , DNA ,