Cloning, Expression and Modification of Antibody Variable Regions (V Regions)

Summary

PCR-based cloning schemes utilize conserved sequences in the framework and pilot regions of the variable region, and although primers can be synthesized with conserved sequences in the framework region for cloning of the variable region, they cause amino acid changes in the framework region, which can affect antigen binding. Therefore, synthesizing primers with conserved hydrophobic pilot sequences is a better solution.

Operation method

Cloning, Expression and Modification of Antibody Variable Region (V Region)

Move

Basic Option 1 Cloning and Expression of Immunoglobulin Variable Regions by Using Redundant Primers

NOTE: Autoclave all instruments and containers that come into contact with RNA and rinse with DEPC-treated water. Experimentation
Wear gloves at all times during the experiment. All water and salt solutions used for RNA are treated with DEPC (Appendix 1).

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