Rapid and simple method for DNA extraction from plant and algal species suitable for PCR amplification using a chelating resin Chelex 100

被引:26
作者
HwangBo, Kwon [1 ]
Son, Su Hyun [1 ,2 ]
Lee, Jong Suk [1 ]
Min, Sung Ran [1 ]
Ko, Suk Min [3 ]
Liu, Jang R. [1 ]
Choi, Dongsu [2 ]
Jeong, Won Joong [1 ]
机构
[1] KRIBB, Plant Syst Engn Res Ctr, Taejon 305806, South Korea
[2] Kunsan Natl Univ, Dept Biol, Gunsan 573701, South Korea
[3] Eugentech Inc, KRIBB, Taejon 305806, South Korea
关键词
Chelex; 100; DNA extraction method; Plant material; PCR; Transgenes; YEAST;
D O I
10.1007/s11816-009-0117-4
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
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.
引用
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页码:49 / 52
页数:4
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