Data and Analysis
LB- Plasmid: LB+ Plasmid: LB/AMP- Plasmid: LB/AMP+ Plasmid:
Prediction: Will have growth Prediction: Will have growth Prediction: Will not have growth Prediction: Will have growth
Reason: No resistance. No Reason: No resistance to Reason: No resistance. Antibiotic Reason: Resistance. Antibiotic
antibiotic. Food for bacteria. antibiotic. Food for bacteria. can kill bacteria. won't work. Food for bacteria.
Observation: Lawn Growth Observation: Lawn Growth Observation: No Growth Observation: 7 colonies
Prediction: Will have growth Prediction: Will have growth Prediction: Will not have growth Prediction: Will have growth
Reason: No resistance. No Reason: No resistance to Reason: No resistance. Antibiotic Reason: Resistance. Antibiotic
antibiotic. Food for bacteria. antibiotic. Food for bacteria. can kill bacteria. won't work. Food for bacteria.
Observation: Lawn Growth Observation: Lawn Growth Observation: No Growth Observation: 7 colonies
Types of Growth
LB- Plasmid: LB+ Plasmid: LB/AMP- Plasmid: LB/AMP+ Plasmid:
~ Positive Control ~ Positive Control ~ Negative Control ~ Experimental
~ Lawn Growth ~ Lawn Growth ~ No Growth ~ 7 Colonies
~ Positive Control ~ Positive Control ~ Negative Control ~ Experimental
~ Lawn Growth ~ Lawn Growth ~ No Growth ~ 7 Colonies
Conclusion Questions
5.
a. LB+ Plasmid and LB- Plasmid both have lawn growth. This shows that bacteria can grow with food present.
b. LB/AMP- Plasmid had no growth while the LB- Plasmid had lawn growth. This shows that the bacteria (LB/AMP-) was killed by the antibiotic,
but if the antibiotic is not present, the LB- Plasmid can grow.
c. LB/AMP- Plasmid had no growth while the LB/AMP+ Plasmid made 7 colonies. This shows that the bacteria (LB/AMP -) was killed by the
antibiotic while the bacteria (LB/AMP+) with the plasmid was able to resist the antibiotic and grow.
d. LB/AMP+ Plasmid produced 7 colonies while the LB+ Plasmid produced lawn growth. This shows that the bacteria (LB/AMP+) took up the
plasmid while the other bacteria (LB+) was weaker and more exposed to the antibiotics effects.
6. In this experiment, antibiotic resistance and how well the plasmid took up the plasmid resistance (the ampicillin worked to allow the bacteria to become resistant to the antibiotics).
7. In this experiment, the phenotype was not observed during observation or incubation time.
8. The plate with the bacteria, plasmid, and antibiotic (LB/AMP+) should be observed to see if the bacteria resists the antibiotics by using the plasmid. If the bacteria is successful, there should be growth.
9.
a. Total Mass of Plasmid Used: .05mg
b. Total Volume of Cell Suspension Prepared: 510 ml
c. Fraction of the Total Cell Suspension That Was Spread on the Plate: 10/51
d. Mass of Plasmid in the Cell Suspension Spread: .0098
e. Number of Colonies in MCG Plasmid DNA: 7.14 x 10^2
10. Some Factors That Might Influence Transformation Efficiency:
~ Amount of Heat Shocking Time - If the bacteria is not heat shocked for an appropriate time, it may not take up the plasmid DNA, and if over heat shocked, the DNA may not even take the DNA at all
~ Growth of Cells in Incubation - If the incubator is not providing an effective constant heat, or if the incubator is too hot/cold, the bacteria may not grow in said environment.
~ DNA Used - If the plasmid DNA happened to be mutated, then the bacteria may not be able to take the plasmid in. It may not be cut properly with the restriction enzyme and may not match the code for the bacteria.
a. LB+ Plasmid and LB- Plasmid both have lawn growth. This shows that bacteria can grow with food present.
b. LB/AMP- Plasmid had no growth while the LB- Plasmid had lawn growth. This shows that the bacteria (LB/AMP-) was killed by the antibiotic,
but if the antibiotic is not present, the LB- Plasmid can grow.
c. LB/AMP- Plasmid had no growth while the LB/AMP+ Plasmid made 7 colonies. This shows that the bacteria (LB/AMP -) was killed by the
antibiotic while the bacteria (LB/AMP+) with the plasmid was able to resist the antibiotic and grow.
d. LB/AMP+ Plasmid produced 7 colonies while the LB+ Plasmid produced lawn growth. This shows that the bacteria (LB/AMP+) took up the
plasmid while the other bacteria (LB+) was weaker and more exposed to the antibiotics effects.
6. In this experiment, antibiotic resistance and how well the plasmid took up the plasmid resistance (the ampicillin worked to allow the bacteria to become resistant to the antibiotics).
7. In this experiment, the phenotype was not observed during observation or incubation time.
8. The plate with the bacteria, plasmid, and antibiotic (LB/AMP+) should be observed to see if the bacteria resists the antibiotics by using the plasmid. If the bacteria is successful, there should be growth.
9.
a. Total Mass of Plasmid Used: .05mg
b. Total Volume of Cell Suspension Prepared: 510 ml
c. Fraction of the Total Cell Suspension That Was Spread on the Plate: 10/51
d. Mass of Plasmid in the Cell Suspension Spread: .0098
e. Number of Colonies in MCG Plasmid DNA: 7.14 x 10^2
10. Some Factors That Might Influence Transformation Efficiency:
~ Amount of Heat Shocking Time - If the bacteria is not heat shocked for an appropriate time, it may not take up the plasmid DNA, and if over heat shocked, the DNA may not even take the DNA at all
~ Growth of Cells in Incubation - If the incubator is not providing an effective constant heat, or if the incubator is too hot/cold, the bacteria may not grow in said environment.
~ DNA Used - If the plasmid DNA happened to be mutated, then the bacteria may not be able to take the plasmid in. It may not be cut properly with the restriction enzyme and may not match the code for the bacteria.