在线客服: 点击这里给我发消息  新用户使用步骤:会员注册→充值→重新登入→进入资源
标题:Flexi Vector Cloning
时间:2021-07-29 09:49:08
DOI:10.1007/978-1-59745-196-3_4
大小:546 kb
页数:20 PAGES
下载: 点击下载
预览:

浏览器不支持嵌入PDF阅读,打开新页面在线阅读

目录:
  • Chapter 4
    • Flexi Vector Cloning
      • 4.1. Introduction
      • 4.2. Materials
        • 4.2.1. Flexi Vector Plasmids
        • 4.2.2. Target Genes
        • 4.2.3. Flexi Vector Reagents
        • 4.2.4. PCR Reagents
        • 4.2.5. Bacterial Cell Culture Materials
        • 4.2.6. Plasmid Preparation
        • 4.2.7. DNA Analysis and Sequence Verification
      • 4.3. Methods
        • 4.3.1. Attachment of Flexi Vector Cloning Sequences
          • 4.3.1.1. PCR Primers
          • 4.3.1.2. PCR Amplification
        • 4.3.2. Restriction Digestion of PCR Products
          • 4.3.2.1. Digestion Reaction
          • 4.3.2.2. Cleanup
        • 4.3.3. Ligation of PCR Products into an Acceptor Vector
        • 4.3.4. Transformation and Growth of Host Cells
        • 4.3.5. Analysis of Plasmid DNA
          • 4.3.5.1. Colony PCR Screening of Transformants
          • 4.3.5.2 Growth of Individual Isolates
          • 4.3.5.3. Qia Prep 96 Turbo Plasmid DNA Purification
          • 4.3.5.4. Determination of DNA Concentration and Purity
          • 4.3.5.5. Sequence Analysis of Plasmid DNA
        • 4.3.6. Creation of Glycerol Stocks
        • 4.3.7. Flexi Vector Transfer Reaction
        • 4.3.8. Alternative Constructs for Flexi Vector
          • 4.3.8.1. Creation of an Antibiotic Resistance Cassette
          • 4.3.8.2. Design of 3′ Sequence in Flexi Vector Plasmids
          • 4.3.8.3. Two-Step PCR for Fusion Protein Expression
          • 4.3.8.4. Native N Terminus
          • 4.3.8.5. C-Terminal Fusion Proteins
      • 4.4. Notes
      • Acknowledgments
      • References

本页内容由网络收集而来,版权归原创者所有,如有侵权请及时联系