从德国小蠊干标本中提取基因组DNA可行性的研究  

Feasibility research of genome DNAs extraction from dried specimen of German cockroach

作　　者：辛娟娟[1] 周小洁[2,3] 李秋红 高俊平[1] 佟颖 曾晓芃[2,3] XIN Juan-juan;ZHOU Xiao-jie;LI Qiu-hong;GAO Jun-ping;Tong Ying;ZENG Xiao-peng(China Agricultural University,Beijing 100193,China;Beijing Center for Disease Control and Prevention,Beijing 100013,China;Beijing Center for Disease Preventive Medical Research,Beijing 100013,China)

机构地区：[1]中国农业大学,北京100193 [2]北京市疾病预防控制中心,北京100013 [3]北京市预防医学研究中心,北京100013

出　　处：《中华卫生杀虫药械》2018年第6期575-577,581,共4页Chinese Journal of Hygienic Insecticides and Equipments

基　　金：国家自然科学基金青年基金项目(编号:31501901);首都卫生发展科研专项项目(编号:2016-4-1011)

摘　　要：目的拟建立用现场采集到的以及饲养过程中德国小蠊死亡后,晾干样本提取基因组DNA的方法。方法从现场环境中采集死亡的、体形较为完整的德国小蠊,晾干后,采用6种预处理方法后进行后续提取,并对所得基因组DNA质量进行吸光度、电泳与PCR扩增检测。结果从现场环境中采集到的德国小蠊干标本中获得了较高质量的基因组DNA,OD_(260/280)值可在1.8~2.0之间,1.5%琼脂糖凝胶电泳在15000bp处有明显条带。以提取到的基因组DNA为模板可扩增出438bp的β-actin基因和超过2000bp的钠离子通道基因VSSC片段。结论以本研究现场环境采集的体形较为完整的德国小蠊干标本为材料可以提取到基因组DNA,能满足至少2000bp的基因片段的PCR扩增要求,该方法在一定条件下有较强的可行性。Objective To establish methods of extracting genome DNAs from dry specimens of German cockroach collected from the field and rearing process.Methods Genome DNA were extracted from dead German cockroach, collected in fields with relatively intact bodies and dried by nature,pre-treated with six different methods.The quality and quantity of DNA were measured through detecting the absorbance,agarose gel electrophoresis,and PCR amplification.Results High quality and quantity DNAs,with OD260/280 values ranged from 1.8 to 2.0 and obvious strips appeared at 15000bp through 1.5% agarose gel electrophoresis,were extracted from the dry specimens of German cockroach sampled in fields.The extracted DNAs met the requirement of PCR amplification for partial of beta- actin gene with 438bp and voltage sensitive sodium channel (vssc)gene more than 2000bp.Conclusion In the present study,relatively intact dry specimens of German cockroach collected in fields were used to extract genome DNA,and the extracted DNA can meet the requirement of PCR amplification for 2000bp at least.This method displays strong feasibility under certain conditions.

关 键 词：德国小蠊 干标本 DNA提取 PCR 

分 类 号：R384.9[医药卫生—医学寄生虫学]