【2017年整理】生物工程实习报告.pptVIP

Novoprotein 专业实践报告 报告人： 学 号： 2014.1.18 Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd. 熟悉、了解、掌握质粒EGFP和LGALS1分别作为模板，设计引物扩增EGFP基因和LGALS1基因，通过Novorec的方法将两个片段同时重组到pET21a载体上的相关理论知识。 绿色荧光蛋白-人LGALS1融合蛋白大肠杆菌的表达载体的构建及纯化 理论学习 Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd. Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd. Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd. 一、 接菌抽提、PCR反应、电泳检测  Kit抽提 ↓ PCR反应 ↓ 电泳上样检测 质粒抽提检测 PCR反应检测 Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd. 二、 酶切过夜、电泳检测、产物回收 PCR反应产物酶切 ↓ 37℃恒温水浴锅过夜 ↓ 配胶 ↓ 电泳检测 ↓ 割胶回收 酶切产物电泳检测 Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd. 三、 重组、转化 、阳性鉴定 目的基因、载体重组 ↓ 转化 ↓ 挑菌、37℃培养 ↓ PCR阳性鉴定 ↓ 电泳检测 Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd. 四、 质粒抽提、电泳检测 碱法抽提质粒→电泳检测 Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd. 五、 SDS电泳检测 Lane A:诱前 Lane B :G-2-1 Lane C:G-2-2 Lane D:Marker Lane E: G-2-3 Lane F:G-2-4 Lane G:G-4-1 Lane H: G-4-2 Lane I : G-4-3 Lane J:G-4-4 A B C D E F G H I J 结论：通过做western blot进一步确定是否为目的条带。 Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd. 六、Western Blot 免疫印迹 标签 C-His 抗体 C-Hi