用于基于PCR技术的高质量基因组DNA的通用快速盐提取法。
Universal and rapid salt-extraction of high quality genomic DNA for PCR-based techniques.
作者信息
Aljanabi S M, Martinez I
机构信息
CENARGEN-EMBRAPA, SAIN-Parque Rural, W5 Norte. C.P. 02372, CEP 70849-970 Brasilia, DF, Brazil.
出版信息
Nucleic Acids Res. 1997 Nov 15;25(22):4692-3. doi: 10.1093/nar/25.22.4692.
Abstract
A very simple, fast, universally applicable and reproducible method to extract high quality megabase genomic DNA from different organisms is described. We applied the same method to extract high quality complex genomic DNA from different tissues (wheat, barley, potato, beans, pear and almond leaves as well as fungi, insects and shrimps' fresh tissue) without any modification. The method does not require expensive and environmentally hazardous reagents and equipment. It can be performed even in low technology laboratories. The amount of tissue required by this method is approximately 50-100 mg. The quantity and the quality of the DNA extracted by this method is high enough to perform hundreds of PCR-based reactions and also to be used in other DNA manipulation techniques such as restriction digestion, Southern blot and cloning.
摘要
本文描述了一种非常简单、快速、普遍适用且可重复的方法，用于从不同生物体中提取高质量的兆碱基基因组DNA。我们应用相同的方法从不同组织（小麦、大麦、马铃薯、豆类、梨和杏仁叶以及真菌、昆虫和虾的新鲜组织）中提取高质量的复杂基因组DNA，无需任何修改。该方法不需要昂贵且对环境有害的试剂和设备。即使在技术水平较低的实验室也可以进行。此方法所需的组织量约为50-100毫克。通过该方法提取的DNA的数量和质量足够高，足以进行数百次基于PCR的反应，也可用于其他DNA操作技术，如限制性酶切、Southern杂交和克隆。
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