Problem
You were provided with the genetic sequence for one of these genes and asked to design a cloning strategy to generate a dsRNA expression construct to knock down that gene by RNAi (pages 4-5).
1. What two restriction sites are you going to use to clone your PCR product into the pL4440 plasmid? What are their DNA sequence?
2. State the primer sequence you will use to amplify the F27C1.7 gene ready to be cloned into the pL4440 plasmid?
3. How would you go about cloning this amplified DNA into pL4440? Using your knowledge of cloning list 5 important aspects of the method.