Tuition Center Singapore JC H2 H1 Biology Tuition – Key Techniques in Genetic Engineering

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Tuition Center Singapore JC H2 H1 Biology Tuition – Key Techniques in Genetic Engineering

1. Insertion of Gene of Interest into a Vector

• Cutting of DNA vector and gene of interest at specific restriction sites by restriction endonucleases (enzymes).
• Annealing of cut vector and gene of interest using DNA ligase, forming recombinant DNA

2. Introduction of Recombinant DNA into Host Organisms.

• Bacterial host cells are induced to take up recombinant DNA by transformation

3. DNA Cloning – A single DNA molecule can be copied to generate many identical molecules by

• Replication of host organism (e.g. bacteria) – in vivo method
• Polymerase chain reaction – in vitro DNA replication process

4. Nucleic Acid Hybridization (not shown in Fig. 1.1)

• technique of using the binding of a complementary nucleic acid sequence (probe) to specific sequence of the gene of interest (DNA or RNA) to confirm the presence of the gene of interest in:

5a. DNA sequencing

• To rapidly sequence all the nucleotides in the gene of interest
• To deduce the amino acid sequence of the protein they encode
• For research purposes

5b. Harvesting Protein Products – Culture the genetically modified bacteria containing the gene of interest in a fermenter on a large scale and harvest the protein products for:

• Commercial use (e.g. Insulin production)
• Basic research on the protein

Please contact Angie @ 96790479 or Mr Ong @ 98639633 if you need tuition in A Level/jC H2 H1 Biology Tuition