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CN101736073A 一种气单胞菌和嗜水气单胞菌双重PCR快速检测试剂盒和检测方法 失效 - 权利终止

一种气单胞菌和嗜水气单胞菌双重PCR快速检测试剂盒和检测方法
摘要：
一种气单胞菌和嗜水气单胞菌双重PCR快速检测试剂盒，包括：(1)DNA提取试剂：TE缓冲液，5-15mM？Tris，0.5-1.5mM？EDTA；10-20mg/mL溶菌酶水溶液；10-20mg/mL蛋白酶K水溶液；Tris饱和酚；2-5mo1/L乙酸钠水溶液；95％乙醇；(2)PCR反应试剂管：PCR反应液49μL，包括10×PCR反应缓冲液5μL，dNTPs(10uM/L)0.5μL，Taq？DNA聚合酶(5U/μL)0.5μL，特异性寡核苷酸引物引物1、引物2、引物3、引物4，用灭菌去离子水定容。一种气单胞菌和嗜水气单胞菌双重PCR快速检测方法：(1)DNA抽提，(2)PCR扩增反应，(3)被检测样品目的PCR扩增产物和电泳检验。本发明适用于水产生物致病菌的快速检测。
摘要（英）：

The invention provides a rapid detection kit of Aeromonas and Aeromonas hydrophila by double PCR, and the kit comprises the following components: (1) DNA extracting reagent: TE buffer solution with 5-15mM of Tris and 0.5-1.5mM of EDTA, lysozyme water solution with the concentration of 10-20mg/mL, protease K water solution with the concentration of 10-20mg/mL, Tris, 2-5mol/L of sodium acetate water solution, and ethanol with the concentration of 95%; (2) PCR reaction reagent pipe: 49mu L of PRC reaction solution comprising 5mu L of 10* PCR reaction buffer solution, 0.5mu L of dNTPs(10mu M/L), 0.5mu L of Taq DNA polymerase (5U/mu L), specific oligonucleotides primer comprising primer 1, primer 2, primer 3 and primer 4 which are in constant volume by sterilized deionized water. The rapid detection method has the following steps: (1) DNA extraction; (2) PCR amplification reaction; and (3) PCR amplification product and electrophoresis detection for detected samples. The invention is applicable to rapid detection of aquatic organism pathogen.

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