MBT FL24 Transformation

Transformation is the insertion of a foreign plasmid into a bacterial cell, resulting in new acquired traits.

1. Bacterial cells are transferred to cold CaCl2. 2. Plasmid with foreign DNA is added to the bacteria. 3. Cells are heat shocked at 42°C. 4. Cells are spread on an agar petri dish. 5. Bacteria are given time to grow on the petri dish.

Ampicillin is used to isolate and select for only the bacteria that have been successfully transformed.

The pGLO LB/amp plate would show no bacterial growth.

The competency of cells is increased through the use of calcium chloride and heat shock.

A plasmid is a small, circular piece of DNA that can replicate independently of chromosomal DNA in bacteria.

Calcium chloride helps to make bacterial cells more permeable to DNA, facilitating the uptake of plasmids.

Heat shock is a brief exposure of bacterial cells to elevated temperatures, which helps to increase the uptake of plasmid DNA.

Spreading transformed bacteria on an agar plate allows for the growth of individual colonies, which can be selected based on antibiotic resistance.

Antibiotic resistance indicates that the bacteria have successfully taken up the plasmid containing the resistance gene, allowing them to survive in the presence of the antibiotic.