高致病性猪繁殖与呼吸综合征病毒湖南株全长感染性cdna克隆的构建

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1、-------国家科技支撑计划资助项目高致病性猪蓝耳病病毒感染性克隆的构建及应用项目编号:2007BAD86B06-4-----------贵州大学硕士学位论文目录目录中文摘要···························································································1ABSTRACT································································

2、······················2前言·································································································4实验研究···························································································51材料······························

3、·········································································51.1毒株、质粒载体与细胞·······································································51.2主要试剂和耗材················································································51.

4、3主要仪器设备···················································································61.4主要溶液的配制················································································72方法···································································

5、··································102.1目的DNA片段的扩增与克隆·····························································102.1.1病毒RNA的制备····································································112.1.2目的DNA片段的RT-PCR扩增·································

6、·················112.1.3PCR产物的电泳与回收纯化······················································13-----------2.1.1PCR产物末端加A与回收纯化···················································132.1.2加A产物与质粒载体的连接······················································142.1

7、.3连接产物的转化······································································142.1.4重组质粒的提取······································································142.1.5重组质粒DNA的定量······························································152.1.6重组质

8、粒的酶切验证与序列测定·················································152.1连接载体的构建··············································································152.2.1质粒载体的提取与定量·····························································152.2.2质