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系统研究核酸沉淀的关键因素,以优化 DNA/RNA 分离。

A systematic investigation of key factors of nucleic acid precipitation toward optimized DNA/RNA isolation.

机构信息

Henan Key Laboratory of Ion-beam Bioengineering, School of Physics, Zhengzhou University, Zhengzhou 450000, China.

Centre for Molecular Medicine & Innovative Therapeutics, Murdoch University, Perth 6150, Australia.

出版信息

Biotechniques. 2020 Apr;68(4):191-199. doi: 10.2144/btn-2019-0109. Epub 2020 Feb 18.

Abstract

Nucleic acid precipitation is important for virtually all molecular biology investigations. However, despite its crucial role, a systematic study of the influence factors of nucleic acid precipitation has not been reported. In the present work, via rational experimental design, key factors of nucleic acid precipitation, including the type of nucleic acid, temperature and time of incubation, speed and time of centrifugation, volume ratio of ethanol/isopropanol to nucleic acid solution, type of cation-containing salt solution and type of coprecipitator, were comprehensively evaluated in an attempt to maximize the efficiency of nucleic acid precipitation. Our results indicate that the optimal conditions of each influence factor of nucleic acid precipitation may vary in accordance with the chemistry, structure and length of nucleic acids.

摘要

核酸沉淀对于几乎所有分子生物学研究都非常重要。然而,尽管它具有关键作用,但目前还没有关于核酸沉淀影响因素的系统研究。在本工作中,通过合理的实验设计,全面评估了核酸沉淀的关键因素,包括核酸类型、孵育温度和时间、离心速度和时间、乙醇/异丙醇与核酸溶液的体积比、含阳离子盐溶液的类型和共沉淀剂的类型,以最大限度地提高核酸沉淀的效率。我们的结果表明,核酸沉淀的每个影响因素的最佳条件可能会根据核酸的化学性质、结构和长度而有所不同。

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