| 固定标本的DNA提取及PCR扩增 |
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| 引用本文： | 高天翔,张秀梅,曾晓起,渡边精一.固定标本的DNA提取及PCR扩增[J].中国海洋大学学报(自然科学版),2000,30(2):244-248. |
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| 作者姓名： | 高天翔 张秀梅 曾晓起 渡边精一 |
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| 作者单位： | 1. 青岛海洋大学水产学院,青岛,266003
2. 日本东京水产大学水生生物系,东京,108-8477 |
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| 基金项目： | 国家自然科学基金课题!(39970 5 78)，日本学术振兴会未来开拓学术研究课题!(97L0 0 90 2 )资助 |
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| 摘 要： | 报道福尔马林、酒精固定的日本绒螯蟹标本的 DNA提取及 PCR扩增 ,并与冷冻、煮沸标本进行了比较。结果表明 ,福尔马林固定标本与酒精固定标本一样可以提取 DNA,加入的蛋白酶 K量越多 ,DNA的回收率越高。以提取的 DNA为模板 ,进行了线粒体 DNA12 S r RNA和 D- loop基因片段的 PCR扩增 ,经琼脂糖凝胶电泳 PCR扩增产物后均得到了与期待的碱基长度相一致的清晰谱带。
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| 关 键 词： | 固定标本 DNA提取 PCR扩增 日本绒螯蟹 |
DNA Extraction and PCR Amplification from Preserved Samples |
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| Gao Tianxiang,Zhang Xiumei,Zeng Xiaoqi,Watanabe Seiichi.DNA Extraction and PCR Amplification from Preserved Samples[J].Periodical of Ocean University of China,2000,30(2):244-248. |
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| Authors: | Gao Tianxiang Zhang Xiumei Zeng Xiaoqi Watanabe Seiichi |
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| Abstract: | DNA extraction and PCR amplification from formalin fixed Japanese mitten crab( Eriocheir japonica De Haan) and alcohol fixed samples were studied and compared with those from frozen samples and boiled samples. The results show that DNA extraction from the formalin fixed samples could be obtained, like that from the alcohol fixed samples. The more the proteinase K we used, the higher the recovery of the DNA was. Using the DNA as a template, we successfully amplified 12S rRNA and D loop region of the mitochondrial DNA by PCR. By agarose gel electrophoresis, we obtained clear bands of the PCR products, whose positions were in accordance with the base pairs expected. |
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| Keywords: | preserved sample DNA extraction PCR amplification Eriocheir japonica |
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