一种改良的CTAB法提取产多糖真菌DNA

doi:10.13523/j.cb.20140510

中国生物工程杂志

2014, Vol. 34

Issue (5): 75-79 DOI: 10.13523/j.cb.20140510

技术与方法

一种改良的CTAB法提取产多糖真菌DNA

刘丽, 张永军, 许长征, 罗锋

西南大学生物能源与环境修复研究中心重庆 400716

A Modified CTAB Method for Isolating Genome DNA from Fungus with Abundant Polysaccharose

LIU Li, ZHANG Yong-jun, XU Chang-zheng, LUO Feng

Research Center of Bioenergy & Bioremediation, Southwest University, Chongqing 400716, China

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摘要：

真菌胞外多糖由于其高吸附高粘稠特点，是困扰从胞外多糖产生菌分离高纯度DNA的难点之一。本文以生产硬葡聚糖的齐整小核菌生产菌为代表，采用改良的CTAB法获得了高质量的基因组DNA。通过分层隔离等培养方法的优化降低硬葡聚糖的产生，并在传统CTAB法的基础上，用高浓度的醋酸钾和无水乙醇共同作用初步沉淀多糖，再用CTAB/NaCl溶液再次去除多糖。相比于商业的DNA提取试剂盒和传统的CTAB法，该方法得到的基因组DNA产率大幅提高，纯度较好，可充分排除胞外多糖的干扰，为各典型产胞外多糖的真菌DNA提取提供重要的参考。提取的基因组DNA可用于基因组文库构建、PCR等分子生物学实验。

关键词： CTAB; DNA提取; 多糖去除

Abstract:

Due to fungal extracellular polysaccharide have high adsorption and high viscous characteristics, it is one of the difficulties of extracting the high purity of nucleic acids from extracellular polysaccharide germs. The production of scleroglucan strain Sclerotium rolfsii as the material, obtained high quality genomic DNA by the improved Cetyltrimethyl Ammonium Bromide (CTAB) method. By reducing the production of polysaccharide in the cell culture process to obtain high mycelium, relatively. And used the high concentration KAC and absolute ethyl alcohol to precipitate the crude polysaccharide, then the CTAB/NaCl solution to remove out the polysaccharide. Compared with the gDNA Kits and traditional method, highly yield and purity of gDNA obtained with simple steps. And the product is suitable for basic molecular experiments, such as building genome libraries and PCR.

Key words: CTAB Extraction Remove out polysaccharide

收稿日期: 2014-02-14 出版日期: 2014-05-25

ZTFLH:

Q89

基金资助:

国家外专干人计划活动专项（2012SWU112097），中央高校基本科研重点项目（XDJK2011B009）资助项目

通讯作者: 罗锋 E-mail: Van77@swu.edu.cn

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引用本文:

刘丽, 张永军, 许长征, 罗锋. 一种改良的CTAB法提取产多糖真菌DNA[J]. 中国生物工程杂志, 2014, 34(5): 75-79.

LIU Li, ZHANG Yong-jun, XU Chang-zheng, LUO Feng. A Modified CTAB Method for Isolating Genome DNA from Fungus with Abundant Polysaccharose. China Biotechnology, 2014, 34(5): 75-79.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20140510 或 https://manu60.magtech.com.cn/biotech/CN/Y2014/V34/I5/75

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