| 不同保存方式下金龟甲虫DNA提取方法及28S rDNA序列分析 |
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| 引用本文： | 贾晨曦,郭晓华,刘广纯.不同保存方式下金龟甲虫DNA提取方法及28S rDNA序列分析[J].沈阳大学学报,2013(6):436-440. |
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| 作者姓名： | 贾晨曦 郭晓华 刘广纯 |
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| 作者单位： | [1]沈阳大学生命科学与工程学院,辽宁沈阳110044 [2]沈阳大学辽宁省城市有害生物治理与生态安全重点实验室,辽宁沈阳110044 |
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| 基金项目： | 辽宁省自然科学基金资助项目（201102157）; 辽宁省科技厅科技计划项目（2008214001） |
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| 摘 要： | 通过电泳检测和PCR扩增28SrDNA核基因序列的结果,比较了无水乙醇、75%乙醇、针插干制3种金龟甲虫标本的保存方式和SDS-蛋白酶K法、CTAB法、饱和NaCl法3种提取DNA方法.结果显示,SDS-蛋白酶K法、CTAB法提取3种保存方式的标本时,可成功提取DNA并有效扩增目的基因;饱和NaCl法仅能提取无水乙醇保存标本的DNA并扩增目的基因,而75%乙醇保存标本和干制标本的DNA提取和基因扩增均不理想.
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| 关 键 词： | 金龟子 标本 保存方法 DNA提取 28S rDNA |
DNA Extraction and 28S rDNA Sequences Analysis of Scarab Beetle Specimens Preserved under Different Conditions |
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| Jia Chenxi,Guo Xiaohua,Liu Guangchun.DNA Extraction and 28S rDNA Sequences Analysis of Scarab Beetle Specimens Preserved under Different Conditions[J].Journal of Shenyang University,2013(6):436-440. |
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| Authors: | Jia Chenxi Guo Xiaohua Liu Guangchun |
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| Institution: | a (a. College of Life Science and Engineering, Shenyang University, b. Liaoning Key Laboratory of Urban Integrated Pest Management and Ecological Security, Shenyang University, Shenyang 110044, China) |
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| Abstract: | The methods of DNA extraction and 28S ribosomal DNA sequences were compared from the specimens of scarab beetle Potosia（L.）brevitarsis（Lewis）which preserved in 100% ethanol,75% ethanol and kept as a dry-pinned specimen.The results show that the genomic DNA were obtained successfully and 28S rDNA sequences were amplified using polymerase chain reaction and sequenced using sequencing techniques from specimens preserved in three conditions by the extraction methods of SDS-protease K and CTAB.However,using the method of saturated NaCl,the specimens that have been fixed in 100% ethanol could yield successful genomic DNA extraction,amplification and sequence information of 28S rDNA sequences,yet preserved dry samples and 75% ethanol fixed samples were quite poor for DNA extraction and 28S rDNA sequences analysis. |
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| Keywords: | scarab beetle specimens preservation methods DNA extraction 28S rDNA |
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