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

PLANT BIOTECHNOLOGY(2008)

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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.
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关键词
Cultivar identification,DNA extraction,DNA marker,glass fiber filter,marker-assist selection
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