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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (34): 12-16.

所属专题： 生物技术； 园艺

• 林学 园艺 园林 • 上一篇    下一篇

巴西橡胶树红根病菌DNA提取有效方法比较

涂敏 蔡海滨 孙爱花 胡彦师   

  • 收稿日期:2012-08-23 修回日期:2012-09-24 出版日期:2012-12-05 发布日期:2012-12-05
  • 基金资助:

    农业部热带作物种植资源保护项目;中国热带农业科学院橡胶研究所所基本科研业务费专项

Comparing Study on Several Methods for DNA Extraction of Ganoderma pseudoferreum

  • Received:2012-08-23 Revised:2012-09-24 Online:2012-12-05 Published:2012-12-05

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摘要：

旨在筛选适用于橡胶树红根病菌DNA提取的有效方法并评价其效果。利用CTAB改良法、氯化苄改良法、SDS改良法等3种方法提取橡胶树红根病菌菌丝体DNA，并通过紫外吸收光谱和琼脂糖凝胶电泳对DNA样品进行检测，比较提取DNA的产量、质量；再选用提取效果最好的方法提取5个时间段（4、6、8、10、12天）的菌丝DNA并进行ITS-PCR检测。CTAB改良法提取的样品DNA纯度最好，产率最高，OD260/OD280为1.82~1.84，浓度可达280 μg/mL；培养4~6天的菌丝提取的DNA质量最高，适合PCR检测需要。CTAB改良法是一种适用于橡胶树红根病菌DNA提取的理想方法。

关键词: 克隆, 克隆

Abstract:

The aim was to screen the suitable DNA extraction method for Ganoderma pseudoferreum of rubber tree and evaluate the effect. The purity and output of Ganoderma pseudoferreum DNA extracted by three improved method including CTAB, benzyl chloride and SDS were compared via ultraviolet absorption spectrum and agarose gel electrophoresis. The best method was used to extract the DNA of five culture time (4, 6, 8, 10, 12 d) and detected by ITS-PCR. The purity and output via improved CTAB showed the best results, OD260/OD280 was 1.82-1.84 and DNA concentration was 280 μg/mL. Cultivating for 4-8 d was the suitable time for DNA extraction and PCR detection. The improved CTAB method was suitable for Ganoderma pseudoferreum DNA extraction.