| DNA recovery from agarose gels with a simple centrifuge-driven sephadex filtration |
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| 作者姓名： | Niu Chen Li Yun |
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| 作者单位： | Niu Chen Li Yun Key Laboratory for Genetics and Breeding in Forest Trees and Ornamental Plants，Ministry of Education，Beijing Forestry University，Beijing 100083，P. R. China Present address: Department of Agronomy and Horticulture，New Mexico State University，Las Cruces，NM 88003，USA |
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| 摘 要： | Conventional methods of DNA recovery from agarose gel generally require expensive equipment, extended elution times, or considerable handling of the sample after elution. We developed a simple protocol for a quick and effective recovery of DNA from agarose gels with good yield and quality. Using a Sephadex resin filled spin column, DNA fragments of 500 bp to 6 kb in an agarose gel slice were easily recovered by a 2 min centrifugation. The recovery efficiencies were over 40%-50% and the eluted DNA can be used directly for downstream application, such as polymerase chain reactions (PCR) and restriction enzyme digestion. This method could also be used to recover large DNA fragment (48 kb) without degradation. The use of Sephadex helps to remove small molecular impurities from agarose and it also reduces the chance of clogging the column filter caused by direct contact with agarose.
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| 关 键 词： | DNA恢复 琼脂糖胶 交联葡聚糖 离心分离机 森林生物学 |
| 收稿时间： | 2005-07-20 |
| 修稿时间： | 2005-10-25 |
DNA recovery from agarose gels with a simple centrifuge-driven Sephadex filtration |
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| Niu Chen Li Yun.DNA recovery from agarose gels with a simple centrifuge-driven sephadex filtration[J].Forestry Studies in China,2006,8(1):32-34. |
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| Authors: | Niu Chen Li Yun |
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| Institution: | 1. Key Laboratory for Genetics and Breeding in Forest Trees and Ornamental Plants, Ministry of Education, Beijing Forestry Univer sity, Beijing 100083, P. R. China;present address: Department of Agronomy and Horticulture, New Mexico State University, Las Cruces, NM 88003, USA
2. Key Laboratory for Genetics and Breeding in Forest Trees and Ornamental Plants, Ministry of Education, Beijing Forestry Univer sity, Beijing 100083, P. R. China |
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| Abstract: | Conventional methods of DNA recovery from agarose gel generally require expensive equipment, extended elution times, or considerable handling of the sample after elution. We developed a simple protocol for a quick and effective recovery of DNA from agarose gels with good yield and quality. Using a Sephadex resin filled spin column, DNA fragments of 500 bp to 6 kb in an agarose gel slice were easily recovered by a 2 min centrifugation. The recovery efficiencies were over 40%-50% and the eluted DNA can be used directly for downstream application, such as polymerase chain reactions (PCR) and restriction enzyme digestion. This method could also be used to recover large DNA fragment (48 kb) without degradation. The use of Sephadex helps to remove small molecular impurities from agarose and it also reduces the chance of clogging the column filter caused by direct contact with agarose. |
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| Keywords: | DNA recovery agarose gel Sephadex filter column centrifuge |
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