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首页> 外文期刊>Journal of Chemical Technology & Biotechnology >A two-buffer method used for both polymerase chain reaction product purification and DNA gel extraction
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A two-buffer method used for both polymerase chain reaction product purification and DNA gel extraction

机译:用于聚合酶链式反应产物纯化和DNA凝胶提取的两缓冲方法

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摘要

BACKGROUND Both polymerase chain reaction (PCR) purification kits and DNA gel extraction kits have been widely used for DNA purification in laboratories worldwide. However, the uses of these kits are costly and produce laboratory wastes that have negative impacts on the environment; it is important to develop a novel method to overcome these drawbacks. RESULTS Here we demonstrate that a saturated sodium iodide solution and a laboratory-made washing buffer can replace the buffers in PCR purification and gel extraction kits for DNA purification and do not affect DNA purification capacity. These two buffers can be utilized to purify DNA along with the columns recycled from PCR purification and DNA gel extraction kits and can maintain DNA purification capacity that is comparable to that of commercial kits. For small-sized DNA fragments, the buffers developed in this study are even more efficient compared to commercially available buffers from PCR purification and DNA gel extraction kits. The quality of DNA purified with these two buffers can meet the requirements of gene cloning. CONCLUSIONS This study provides a two-buffer method that is used for both PCR product purification and DNA gel extraction. This method is simple, of low cost and beneficial to numerous laboratories worldwide. (c) 2019 Society of Chemical Industry
机译:背景技术聚合酶链反应(PCR)纯化试剂盒和DNA凝胶提取试剂盒已广泛用于全球实验室中的DNA净化。然而,这些套件的用途昂贵,生产对环境产生负面影响的实验室废物;开发一种克服这些缺点的新方法非常重要。结果我们证明饱和碘化钠溶液和实验室制备的洗涤缓冲液可以代替PCR纯化和凝胶提取试剂盒中的缓冲剂,用于DNA纯化,不会影响DNA净化能力。这两个缓冲液可用于纯化DNA以及从PCR纯化和DNA凝胶提取试剂盒中再循环的柱,并且可以维持与商业套件相当的DNA净化能力。对于小尺寸的DNA片段,与来自PCR纯化和DNA凝胶提取试剂盒的市售缓冲液相比,该研究中开发的缓冲剂更有效。用这两个缓冲液纯化的DNA质量可以满足基因克隆的要求。结论本研究提供了一种双缓冲方法,用于PCR产物纯化和DNA凝胶萃取。这种方法很简单,成本低,有利于全球众多实验室。 (c)2019年化学工业协会

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