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首页> 外文期刊>Plant Biotechnology Reports >Rapid and simple method for DNA extraction from plant and algal species suitable for PCR amplification using a chelating resin Chelex 100
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Rapid and simple method for DNA extraction from plant and algal species suitable for PCR amplification using a chelating resin Chelex 100

机译:使用螯合树脂Chelex 100从植物和藻类物种中提取DNA的快速简便方法,适用于PCR扩增

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摘要

A DNA extraction method using Chelex 100 is widely used for bacteria, Chlamydomonas, and animal cell lines, but only rarely for plant materials due to the need for additional time-consuming and tedious steps. We have modified the Chelex 100 protocol and successfully developed a rapid and simple method of DNA extraction for efficient PCR-based detection of transgenes from a variety of transgenic plant and algal species. Our protocol consists of homogenizing plant tissue with a pestle, boiling the homogenized tissue in a microfuge tube with 5% Chelex 100 for 5 min, and centrifuging the boiled mixture. The supernatant, which is used for PCR analysis, was able to successfully amplify transgenes in transgenic tobacco, tomato, potato, Arabidopsis, rice, strawberry, Spirodela polyrhiza, Chlamydomonas, and Porphyra tenera. The entire DNA extraction procedure requires <15 min and is therefore comparable to that used for bacteria, Chlamydomonas, and animal cell lines. Keywords Chelex 100 - DNA extraction method - Plant material - PCR - Transgenes
机译:使用Chelex 100的DNA提取方法被广泛用于细菌,衣原体和动物细胞系,但由于需要额外的费时且繁琐的步骤,因此很少用于植物材料。我们已经修改了Chelex 100协议,并成功开发了一种快速,简单的DNA提取方法,用于基于PCR的高效检测各种转基因植物和藻类中的转基因。我们的方案包括用杵将植物组织匀浆,在装有5%Chelex 100的微量离心管中将匀浆的组织煮沸5分钟,然后将煮沸的混合物离心。用于PCR分析的上清液能够成功扩增转基因烟草,番茄,马铃薯,拟南芥,水稻,草莓,螺旋藻,衣藻和紫菜中的转基因。整个DNA提取过程需要不到15分钟,因此与细菌,衣藻和动物细胞系所用的DNA相当。关键词Chelex 100-DNA提取方法-植物材料-PCR-转基因

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