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应用于药用蘑菇灵芝属PCR鉴定的快速DNA提取方法比较

Comparison of rapid DNA extraction methods applied to PCR identification of medicinal mushroom Ganoderma spp.

作者信息

Zhou Xuanwei, Li Qizhang, Zhao Jingya, Tang Kexuan, Lin Juan, Yin Yizhou

机构信息

Plant Biotechnology Research Center, School of Agriculture and Biology, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center Shanghai Jiao Tong University, Shanghai, PR China.

出版信息

Prep Biochem Biotechnol. 2007;37(4):369-80. doi: 10.1080/10826060701593282.

Abstract

Four different DNA extraction methods were used to extract genomic DNA of the medicinal mushroom Lingzhi from its developing stage materials, such as mycelium, dry fruiting body, or sliced and spore powder or sporoderm-broken spore powder. The DNA samples were analyzed using agarose gel electrophoresis, UV spectrophotometer, and PCR amplification. According to the average yields and purity of DNA, high salt concentrations and low pH methods were the best for DNA extraction. The mycelia and sporoderm-broken spore powder yielded higher and purer DNA. The method developed could effectively eliminate the influence of the secondary metabolites to DNA extraction. The DNA samples extracted from the developed method could be successfully used for PCR applications.

摘要

采用四种不同的DNA提取方法,从灵芝发育阶段的材料(如菌丝体、干子实体、切片及孢子粉或破壁孢子粉)中提取其基因组DNA。使用琼脂糖凝胶电泳、紫外分光光度计和PCR扩增对DNA样本进行分析。根据DNA的平均产量和纯度,高盐浓度法和低pH法最适合DNA提取。菌丝体和破壁孢子粉产生的DNA产量更高、纯度更高。所开发的方法可以有效消除次生代谢产物对DNA提取的影响。从所开发方法中提取的DNA样本可成功用于PCR应用。

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