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巴西橡胶树红根病菌DNA提取有效方法比较
引用本文:涂敏,蔡海滨,孙爱花,胡彦师.巴西橡胶树红根病菌DNA提取有效方法比较[J].中国农学通报,2012,28(34):12-16.
作者姓名:涂敏  蔡海滨  孙爱花  胡彦师
作者单位:中国热带农业科学院橡胶研究所/农业部橡胶树生物学与遗传资源利用重点实验室/国家橡胶树育种中心,海南儋州,571737
基金项目:农业部热带作物种植资源保护项目"橡胶树种质资源保护",中国热带农业科学院橡胶研究所所基本科研业务费专项"橡胶树红根病菌毒素对橡胶树无性系砧木的抗性筛选和抗性诱导研究"
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
收稿时间:2012/8/23 0:00:00
修稿时间:2012/9/24 0:00:00

Comparing Study on Several Methods for DNA Extraction of Ganoderma pseudoferreum
Tu Min , Cai Haibin , Sun Aihua , Hu Yanshi , Hua Yuwei , Huang Huasun.Comparing Study on Several Methods for DNA Extraction of Ganoderma pseudoferreum[J].Chinese Agricultural Science Bulletin,2012,28(34):12-16.
Authors:Tu Min  Cai Haibin  Sun Aihua  Hu Yanshi  Hua Yuwei  Huang Huasun
Institution:(Ministry of Agriculture Key Laboratory of Biology and Genetic Resources of Rubber Tree/State Center for Rubber Tree Breeding/Rubber Research Institute,CATAS,Danzhou Hainan 571737)
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.

Keywords:

Improved CTAB method

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