Positive E. Coli Lab Report
Molecular Biology
Lab Report
Payton Jackson
Introduction
In this lab, I am going to use antibiotic-resistance plasmids to transform Escherichia coli.
Materials
For this lab you will need the following:
LB Agar
Petri dishes
Beakers
Test tubes
CaCl2 solution
Sensitive E. coli (-ampR) amp plasmids ampicillin -amp cells
Water bath to heat shock cells
A freezer to incubate cells
Process
Step 1: Wash hands and sanitize lab setting. This will prevent anything reacting with a substance that could have been present when it shouldn’t have been.
Step 2: Ampicillin sensitive E. coli cells in log phase of growth are transferred to cold CaCl2 solution.
Step 3: ampR plasmids are added to experimental cells only.
Step 4: Cells are heat-shocked at 42oC. Some of the competent cells take up the ampR plasmid and are transformed.
Step 5: The treated cells are spread on an agar plate containing ampicillin. Ampicillin …show more content…
This is when a host organism takes in foreign DNA and expresses the foreign gene. We used E. coli in this lab because it grows very rapidly. We used plasmids to enter the foreign DNA into the cells. We then made our cells competent by a process that uses calcium chloride and heat shock. We learned that there is no ampicillin in the LB agar because E. coli, which is sensitive to the ampicillin, covered the plate with a lawn of cells. The reason we used LB (Luria Broth) is because it is food, and it will help the bacteria grow. We added ampR plasmids to the experimental cells and heat-shocked them. When the cells were spread on the LB agar containing ampicillin and incubated for 24 hours, it showed the cells transformed. Only transformed cells can grow on agar with ampicillin. We know that the ampicillin was present because there were no lawns present. If the ampicillin, or antibiotic, wasn’t present, the E. coli would have just grown everywhere instead of in
Lab Report
Payton Jackson
Introduction
In this lab, I am going to use antibiotic-resistance plasmids to transform Escherichia coli.
Materials
For this lab you will need the following:
LB Agar
Petri dishes
Beakers
Test tubes
CaCl2 solution
Sensitive E. coli (-ampR) amp plasmids ampicillin -amp cells
Water bath to heat shock cells
A freezer to incubate cells
Process
Step 1: Wash hands and sanitize lab setting. This will prevent anything reacting with a substance that could have been present when it shouldn’t have been.
Step 2: Ampicillin sensitive E. coli cells in log phase of growth are transferred to cold CaCl2 solution.
Step 3: ampR plasmids are added to experimental cells only.
Step 4: Cells are heat-shocked at 42oC. Some of the competent cells take up the ampR plasmid and are transformed.
Step 5: The treated cells are spread on an agar plate containing ampicillin. Ampicillin …show more content…
This is when a host organism takes in foreign DNA and expresses the foreign gene. We used E. coli in this lab because it grows very rapidly. We used plasmids to enter the foreign DNA into the cells. We then made our cells competent by a process that uses calcium chloride and heat shock. We learned that there is no ampicillin in the LB agar because E. coli, which is sensitive to the ampicillin, covered the plate with a lawn of cells. The reason we used LB (Luria Broth) is because it is food, and it will help the bacteria grow. We added ampR plasmids to the experimental cells and heat-shocked them. When the cells were spread on the LB agar containing ampicillin and incubated for 24 hours, it showed the cells transformed. Only transformed cells can grow on agar with ampicillin. We know that the ampicillin was present because there were no lawns present. If the ampicillin, or antibiotic, wasn’t present, the E. coli would have just grown everywhere instead of in