论文部分内容阅读
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 ,用于 PCR检测 HHNBV能保证较高的灵敏度和专一性
Chinese shrimp tissues artificially infected with HHNBV (an isolate of WSSV) were maintained using a sampling solution SEMP-Tris and then purified by phenol extraction, glass milk recovery, nitrocellulose membrane binding and boiling-ethanol precipitation Method to extract DNA. The extracted DNA was amplified by PCR using HHNBV primer, and the amplified product was identified and detected by agarose gel electrophoresis. By comparison, the boiled-ethanol precipitation method is the most rapid and simple method for preparing PCR template from the shrimp tissue preserved by SEMP-Tris in the sample liquid. The PCR method for PCR detection of HHNBV can ensure high sensitivity and specificity