Rapid and simple method for DNA extraction from plant and algal species suitable for PCR amplification using a chelating resin Chelex 100
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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DOI：
10.1007/s11816-009-0117-4
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发表时间：
2010-01-01
影响因子：
2.4
通讯作者：
Jeong, Won Joong
中科院分区：
文献类型：
--
作者：
HwangBo, Kwon;Son, Su Hyun;Jeong, Won Joong
使用CHELEX 100的DNA提取方法被广泛用于细菌、衣藻和动物细胞系，但很少用于植物材料，因为需要额外的耗时和繁琐的步骤。我们已经修改了Cherex 100协议，并成功地开发了一种快速而简单的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物种的转基因。我们的程序包括用筷子将植物组织均质，在微离聚管中用5%的CHELEX 100煮沸5分钟，然后将煮沸的混合物离心。该上清液可成功地扩增出转基因烟草、番茄、马铃薯、拟南芥、水稻、草莓、紫菜、衣藻和紫菜的转基因片段。整个DNA提取过程需要15分钟，因此可以与细菌、衣藻和动物细胞系的提取过程相媲美。
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.