Post-Lab Questions:
1. Exogenous DNA does not passively enter E. coli cells that are not competent. What treatment do cells require to be competent?
The cells are given DNA in the presence of calcium, rubidium, or magnesium chloride and the suspension is moved from one temperature to another very quickly. That makes the cell wall and membrane permeable so the DNA can enter the cell.
2. Why doesn’t the recovery broth used in this experiment contain ampicillin?
Both the -DNA and the +DNA microfuge get recovery broth and if the the -DNA gets the Recovery broth it would kill the E. coli there which would not show how the DNA transforms the bacteria in the +DNA plates.
3. What evidence do you have that transformation was successful?
The cells were not killed by the ampicillin in the +DNA containers but in the -DNA/+AMP so the transformation was successful because the cells that were meant to be transformed showed the desired results of growth and the non-transformed cells did not grow but died.
4. What are some reasons why transformation may not be successful?
If the condition of changing the temperature to help the transformation along were not meant fully the cells would not become vulnerable to the DNA so that they could transform.
5. What is the source of the fluorescence? Why are some cells fluorescent and other cells not fluorescent?
The transformed cells with DNA added because they are made to contain Green Fluorescent Protein so it absorbs blue light and gives off green light as a response.
1. Exogenous DNA does not passively enter E. coli cells that are not competent. What treatment do cells require to be competent?
The cells are given DNA in the presence of calcium, rubidium, or magnesium chloride and the suspension is moved from one temperature to another very quickly. That makes the cell wall and membrane permeable so the DNA can enter the cell.
2. Why doesn’t the recovery broth used in this experiment contain ampicillin?
Both the -DNA and the +DNA microfuge get recovery broth and if the the -DNA gets the Recovery broth it would kill the E. coli there which would not show how the DNA transforms the bacteria in the +DNA plates.
3. What evidence do you have that transformation was successful?
The cells were not killed by the ampicillin in the +DNA containers but in the -DNA/+AMP so the transformation was successful because the cells that were meant to be transformed showed the desired results of growth and the non-transformed cells did not grow but died.
4. What are some reasons why transformation may not be successful?
If the condition of changing the temperature to help the transformation along were not meant fully the cells would not become vulnerable to the DNA so that they could transform.
5. What is the source of the fluorescence? Why are some cells fluorescent and other cells not fluorescent?
The transformed cells with DNA added because they are made to contain Green Fluorescent Protein so it absorbs blue light and gives off green light as a response.
*For some of the things that might be harder to see in the infographic.*
Procedures
Conclusion
Pre-Lab Questions:
Transformation Energy:
Procedures
Conclusion
Pre-Lab Questions:
Transformation Energy:
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