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ABOUT THE VIRTUAL LAB SIMULATION

In the Synthetic Biology Lab, students will learn all the steps involved in constructing and testing a genetic biological circuit. This first lab focuses on the the many steps involved to clone a biological circuit and multiply it in bacterial cells.

SUMMARIZED LEARNING OUTCOMES

- Learn how to engineer natural systems to perform specific functions.
- Learn the fundamentals of the Gateway cloning technique and design your own biological circuit.
- Understand and perform bacterial transformation, antibiotic selection and plasmid purification.
- Understand and perform a restriction digest of your cloning product.

DETAILED LAB DESCRIPTION

Design a biological circuit

Use the specific microRNA profile of cancer cells to design an apoptotic biological circuit that is only activated in cancer cells. You are the only hope for a patient with a rare form of cancer.

Gateway cloning and transformation

Use the Gateway cloning technique to combine different genetic modules in an expression vector. Learn how the Gateway cloning technique can be used to efficiently combine these modules. Use electroporation to transform the bacteria with your vector of interest and select successful transformants. You will learn the basics of sterile lab work and bacterial selection.

Plasmid purification and mapping

Grow the transformed bacteria and extract the plasmids using a ‘miniprep’ kit. Learn how different buffers enable you to isolate plasmid, and not genomic, DNA from the cells. Check if the plasmid, indeed, contains your circuit, and if it is mutation-free by digesting it with restriction enzymes and by using gel electrophoresis.

Test your circuit

In the end you will test your circuit in living cells to see if the cancer cells, indeed, are the only cells that die. Will you be able to find a cure for this rare form of cancer?

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