Answer to Question #190940 in Molecular Biology for Munmun

Question

Describe how you can clone the gene encoding the protein R in the vector pQE-70. Your description should also indicate the choice of restriction enzymes that can be used, the process of screening of E.coli cells harbouring the recombinant plasmid.

Answer 1

Experimental Procedure

• Do a genomic DNA extraction
• Run PCR and purify the PCR product: Run PCR to amplify your insert DNA.
• Digest your DNA:
• Isolate your insert and vector by gel purification:
• Ligate your insert into your vector: DNA ligation and transformation of the plasmid DNA into E.coli.
• Isolate the Finished Plasmid:
• Verify your Plasmid by Sequencing: DNA sequencing