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药用植物艾纳香基因组DNA提取方法研究 |
庞玉新1,2, 王文全1*, 张影波2,3, 刘国道2, 莫廷辉3
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1.北京中医药大学 中药学院, 北京 100102;2.中国热带农业科学院 热带作物品种
资源研究所, 海南 儋州 571737;3.海南大学 农学院, 海南 儋州 571737
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摘要: |
以药用植物艾纳香为研究对象,以-20 ℃保存、4 ℃保存、室温自然干燥和硅胶干燥四种样品保存方式,并采用SDS法、CTAB法、SDS-CTAB法和改良CTAB法4种不同的基因组DNA提取方法进行了对比试验,以期建立艾纳香的较好的样品保存方法和基因组DNA提取方法。结果表明,-20 ℃保存是艾纳香的较理想的样品保存方式; 改良CTAB法是艾纳香基因组DNA提取较适宜的方法,该方法提取的DNA经紫外检测,其A260/A280为1.8左右,明显优于SDS法(1.1~1.5)、CTAB法(1.2~1.5)和SDS-CTAB法(1.4~1.6),琼脂糖凝胶电泳、酶切检测和PCR扩增也得出了同样的结论。 |
关键词: 艾纳香 基因组DNA 提取 方法 |
DOI: |
分类号:Q943 |
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Method of genomic DNA extraction of the herb Blumea balsamifera |
PANG Yu-Xin1,2, WANG Wen-Quan1*, ZHANG Ying-Bo2,3,
LIU Guo-Dao 2, MO Ting-Hui3
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1.College of Chinese Medical Sciences, Beijing University of Traditional Chinese Medicine, Beijing 100102, China;2.China Tropical Crops Genetic Resources Institute of Chinese Academy of Tropical Agricultural Sciences,
Danzhou 571737, China;3.College of Agriculture, Hainan University, Danzhou 571737, China
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Abstract: |
The paper dealt with setting up a more suitable sample preservation way and genomic DNA extraction method for Blumea balsamifera by comparing and analysing four sample preservation ways(preserved at -20 ℃,preserved at 4 ℃,natural drying and silica gel drying)and extraction methods(SDS,CTAB,SDS-CTAB,and modified of CTAB). The results showed that -20 ℃ was a better way for sample preservation and the modified CTAB method was a better method for DNA extraction than others. The UV detection value of A260/A280 was 1.7-1.8 for DNA extracted by this modified CTAB method,obviously higher than that of others,such as 1.1-1.5 for SDS method,1.2-1.5 for CTAB method,1.4-1.6 for SDS-CTAB method. The agarose electrophoresis,restriction endonuclease digestion detection and PCR amplification also indicated that the modified of CTAB method was better than others. |
Key words: Blumea balsamifera genomic DNA extraction method |