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用于PCR检测对虾皮下及造血组织坏死杆状病毒(HHNBV)的一种快速提取DNA方法
引用本文:王馗,史成银,黄倢.用于PCR检测对虾皮下及造血组织坏死杆状病毒(HHNBV)的一种快速提取DNA方法[J].海洋水产研究,2000,21(1):8-12.
作者姓名:王馗  史成银  黄倢
作者单位:1. 青岛海洋大学生命学院,青岛,266003
2. 中国水产科学研究院黄海水产研究所,青岛,266071
基金项目:国家“九五”攻关项目(9600503102)资助
摘    要:使用采样液SEMP-Tris保存人工感染HHNBV(WSSV的一个分离株)的中国对虾组织,然后分别用酚抽提法、玻璃乳(Glass Milk)回收法、硝酸纤维素膜结合法和煮沸-乙醇沉淀法提取DNA。应用HHNBV引物对提取的DNA进行PCR扩增,使用琼脂糖凝胶电泳对扩增产物进行鉴定和检测。通过比较表明,煮沸-乙醇沉淀法是一种最快速、简便的从采样液SEMP-Tris保存的病虾组织中制备PCR模板的方法

关 键 词:模板制备  HHNBV  对虾病毒  DNA提取法  PCR检测

THE COMPARISION OF DIFFERENT METHODS OF DNA EXTRACTING APPLIED IN PCR DETECTION OF HYPODERMAL AND HEMATOPOIETIC NECROSIS BACULOVIRUS
Wang Kui,Shi ChengYin,Huang Jie.THE COMPARISION OF DIFFERENT METHODS OF DNA EXTRACTING APPLIED IN PCR DETECTION OF HYPODERMAL AND HEMATOPOIETIC NECROSIS BACULOVIRUS[J].Marine Fisheries Research,2000,21(1):8-12.
Authors:Wang Kui  Shi ChengYin  Huang Jie
Institution:Wang Kui ,Shi ChengYin ,Huang Jie ;(Yellow Sea Fisheries Research Institute,Qingdao 266071)
Abstract:The tissues of Penaeus Chinensis artificially infected with hypodermal and hematopoietic neccrosis baculovirus (HHNBV, isolated strain of WSSV) were stored in SEMP Tris. DNA were extracted by applying the methods of phenol chloroform extraction, glass milk extraction, nitrocellulose membrane (NCM) adsorption and boiling ethanol precipitation respectirely. After PCR amplifying, the PCR products were detected by agarose electrophoresis. By comparison, the extracting method of boiling ethanol precipitation was considered as a quick, simple method for the preparation of PCR template from the SEMP stored tissues. It can guarantee the high sensitivity and specification of PCR of HHNBV.
Keywords:Preparation of PCR template  HHNBV (isolated strain of WSSV)  PCR
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