Materials Required

1. Petri Dishes
2. Test Tube
3. Restriction Enzyme
4. Vector
5. Plant
6. Nutrient Agar

Procedure

1. Extract the DNA from the organism containing the gene of interest.
2. Use specific restriction enzymes (restriction enzyme digestion or PCR) to cut the DNA into fragments, including the desired gene.
3. Take a Petri dish containing bacterial plasmids (circular DNA molecules) that will serve as vectors.
4. Mix the cut DNA fragments with the plasmids. The gene of interest is inserted into the plasmid using the same restriction enzyme, followed by the enzyme DNA ligase to seal the gaps, forming recombinant DNA.
5. The result is a test tube containing recombinant DNA molecules (plasmids carrying the gene of interest).
6. Introduce the recombinant plasmids into competent bacterial cells, usually by heat shock or electroporation.
7. Plate the transformed bacteria onto a nutrient agar medium. Allow the bacteria to grow and form colonies.
8. Identify colonies that contain the gene of interest by one or more methods: - Checking the DNA sequence of the insert - Detecting the protein product encoded by the gene - Using a DNA marker closely linked to the gene of interest