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有效抽提乙醇中螨标本的核DNA

EFFICIENT EXTRACTION OF DNA FROM ALCOHOL-PRESERVED SPIDER MITES

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【作者】 董慧琴刁兆彦杨琰云陈燃

【Author】 Dong Huiqin,Diao Zhaoyan,Yang Yanyun and Chen Ran(School of Life Science ,Fudan University Shangai 200433)

【机构】 复旦大学生命科学学院复旦大学生命科学学院 上海 200433上海 200433上海 200433

【摘要】 对如何有效地从乙醇保存的叶螨中抽提核基因组DNA作了探讨。70%乙醇保存的叶螨标本,经过干燥,TEN洗涤,蛋白酶K消化,酚—氯仿抽提,2倍体积无水乙醇沉淀等步骤,得到叶螨核基因组DNA沉淀,再用TE或无菌水溶解后,以其为模板,进行随机引物PCR扩增。结果显示.该方法抽提的DNA无Taq酶抑制物,且其纯度达到进行PCR反应对模板要求的程度。这为采用AP-PCR技术研究螨的系统分类提供了便利。

【Abstract】 A method have been developed to extract DNA from alcohol-preserved spider miles (Acari:Tetrynuchus cinnabarinus Biosduval). Alcohol must be evaporated from each specimen by being picked up on the filter paper. Then debris from the mites was removed in TEN buffer by 2-3 squential washing steps. To monitor the efficiency of this method,a polymerase chain reaction (PCR) was designed to amplify DNA of T. cinnabarinus. Detection of amplification products from the PCR indicated that DNA had been successfully extracted and that Taq-polymerase inhibitors were absent. This method was a useful tool in systematic taxnomy of spiter mites and the further analysis research for others.

【关键词】 叶螨DNA抽提AP-PCR
【Key words】 Mites DNA extraction AP-PCR
【基金】 高等学校博士学科点专项科研基金
  • 【文献出处】 蛛形学报 ,Acta Arachnologica Sinica , 编辑部邮箱 ,1998年01期
  • 【分类号】Q953
  • 【被引频次】3
  • 【下载频次】47
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