Rapid and simple DNA extraction method from rice using a glass-fiber filter inserted pipette tip

被引:3
|
作者
Fukami, Masanobu [1 ]
Muramoto, Yasunori [2 ]
Ohkoshi, Kazuo [1 ]
机构
[1] Chiba Prefectural Agr & Forestry Res Ctr, Chiba 2660006, Japan
[2] Gifu Prefectural Agr Technor Ctr, Gifu 5011152, Japan
关键词
Cultivar identification; DNA extraction; DNA marker; glass fiber filter; marker-assist selection;
D O I
10.5511/plantbiotechnology.25.493
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 novel and simple DNA extraction method was developed, which was called "the filter-inserted tip method". This method used glass fiber filters inserted within micropipette tips. It was conducted by drawing and expelling fluid by pipetting using a multichannel pipette. This allowed us to manipulate a large number of samples at the same time for DNA adsorption, fixing, washing and extraction. The 3mm x 3 mm size of glass fiber filter was suitable for this method. To examine the reproducibility of the method, DNA extraction experiment was carried out using rice leaves, and the yielded DNA samples were used for PCR reactions. As a result, PCR amplifications were recognized for 234 out of 236 samples examined, showing high reproducibility of the method. It took approximately 20 min to conduct DNA extractions from 16 samples. These results suggest that this method is suitable for rapid DNA extractions from a large number of samples for PCR reactions, such as marker-assisted selection with simple manipulations, low cost, and without using expensive equipment.
引用
收藏
页码:493 / 496
页数:4
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