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非酶盐析法从酵母和其他真菌中提取改良的酚/氯仿免费 DNA。

Modified Phenol/Chloroform-Free DNA Isolation from Yeast and Other Fungi by Non-Enzymatic Salting Out Method.

机构信息

Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

出版信息

Curr Protoc. 2023 Apr;3(4):e749. doi: 10.1002/cpz1.749.

Abstract

The most commonly used fungal DNA extraction techniques require enzymatic digestion or hazardous chemicals. Here, we describe a quick non-enzymatic salt-out method that we developed for isolating high-quality DNA from various fungal species. DNA was extracted from four species of typical fungi, including Saccharomyces cerevisiae, Hansenula polymorpha, Candida albicans, and Aspergillus flavus. Results that we obtained from agarose gel electrophoresis and spectrophotometry verified the integrity, high quality, and purity of the extracted DNAs. Also, PCR amplification and restriction enzyme digestion confirmed the efficiency of the extracted DNAs in molecular biology applications. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Yeast and fungal cultivation for DNA extraction Basic Protocol 2: Modified salting out DNA extraction protocol Support Protocol 1: PCR amplification of extracted genomes Support Protocol 2: Digestion with restriction enzyme.

摘要

最常用的真菌 DNA 提取技术需要酶消化或危险化学品。在这里,我们描述了一种快速的非酶盐析方法,用于从各种真菌物种中分离高质量的 DNA。从四种典型真菌(包括酿酒酵母、多形汉逊酵母、白色念珠菌和黄曲霉)中提取了 DNA。琼脂糖凝胶电泳和分光光度法的结果验证了提取 DNA 的完整性、高质量和纯度。此外,PCR 扩增和限制性内切酶消化证实了提取 DNA 在分子生物学应用中的效率。© 2023 威利父子公司。基本方案 1:用于 DNA 提取的酵母和真菌培养基本方案 2:改良盐析 DNA 提取方案支持方案 1:提取基因组的 PCR 扩增支持方案 2:用限制酶消化。

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