Dna Transformation Lab
Conclusions of DH5α DNA transformation with red colonies resistance to ampicillin and the lacZ gene
Introduction:
In this experiment, a plasmid with a gene that has resistance to the antibiotic ampicillin and has lacZ is used to transfer the resistance into E. coli bacteria in red colonies. This same technique is used to give diabetics their insulin, and to give dwarfs growth hormones. The point of this lab is to give the groups an idea how DNA can be transformed by a bacteria to improve the lives of people. Transformation happened when a gene is transferred from one bacterium to another one on a plasmid. E. coli is the organism that genetics prefer to use because it can be easily grown and suspended …show more content…
* DH5α (E. coli). * Plasmid DNA . * LB Broth. * LB ager. * 2 microcentrifuge tubes (1.5 ml) containing 2 drops of sterile CaCl2 and labeled "CaCl2". * 4 sterile plastic pipettes. * 1 aluminum foil packet containing 4 sterile paper clips that are large and smooth. The clips should be opened into a 90o angle and the small end bent to close it. * 1 Sharpie marking pen. * 1 glass test tube with a cap containing 2 ml of sterile nutrient broth and labeled "Broth". * 2 Petri dishes containing only nutrient agar and labeled "No Amp" on the bottom with date. * 2 Petri dishes containing nutrient agar and the antibiotic ampicillin. The dishes should be labeled "Amp" on the bottom with the date. * The laboratory instructions.
Methods: Pre-Lab: * PREPARATION OF THE E. COLI STARTER PLATE * One petri dish containing live DH5α E. coli. Use a sterilized transfer loop, a paper clip bent into a loop and sterilized, or a sterilized toothpick. Use the device to touch a colony of bacteria from a petri dish or test tube. Spread the bacteria on the plate in a zig-zag pattern to obtain individual colonies as the concentration of bacteria on the transfer device becomes less. Incubate the plates at 37o C for 24-36 hours. Colonies should grow to the size of this 0 for use in the lab …show more content…
Results: (For the group of this lab report)
Petri dish 1- LB and B1- Lawn growth all over the dish
Petri dish 2- LB AMP and B1-
Petri dish 3- LB and B2- Lawn growth all over the dish
Petri dish 4- LB AMP/ Mac and B2 (control) – showed no signs of grown at all
Petri Dish 1- lawn growth all over Mac B1 Petri Dish 2- Mac/ Amp B1 350 Colonies
Petri Dish 3- Lawn growth Mac B2 Petri Dish 4- Control Mac/ Amp no growth
Conclusion:
All of the groups in the class got the correct results, which means, no one had any contaminations. Some groups had more than colonies on Petri dish 2, and that just depends on how much the drops that were put on or grew. When we did the drops, not all of the drops grew because only certain plasmids took on the gene for resistance. When you have lawn growth on a plate, it means there are no obvious colonies, because they are all over on the plate. Petri dish 1 and 3 were lawn growth just as expedited. Petri dish 4 was our control and had no growth on it that the group could tell; which is what was expedited. All groups had no growth in the control plate; which once again means no
Introduction:
In this experiment, a plasmid with a gene that has resistance to the antibiotic ampicillin and has lacZ is used to transfer the resistance into E. coli bacteria in red colonies. This same technique is used to give diabetics their insulin, and to give dwarfs growth hormones. The point of this lab is to give the groups an idea how DNA can be transformed by a bacteria to improve the lives of people. Transformation happened when a gene is transferred from one bacterium to another one on a plasmid. E. coli is the organism that genetics prefer to use because it can be easily grown and suspended …show more content…
* DH5α (E. coli). * Plasmid DNA . * LB Broth. * LB ager. * 2 microcentrifuge tubes (1.5 ml) containing 2 drops of sterile CaCl2 and labeled "CaCl2". * 4 sterile plastic pipettes. * 1 aluminum foil packet containing 4 sterile paper clips that are large and smooth. The clips should be opened into a 90o angle and the small end bent to close it. * 1 Sharpie marking pen. * 1 glass test tube with a cap containing 2 ml of sterile nutrient broth and labeled "Broth". * 2 Petri dishes containing only nutrient agar and labeled "No Amp" on the bottom with date. * 2 Petri dishes containing nutrient agar and the antibiotic ampicillin. The dishes should be labeled "Amp" on the bottom with the date. * The laboratory instructions.
Methods: Pre-Lab: * PREPARATION OF THE E. COLI STARTER PLATE * One petri dish containing live DH5α E. coli. Use a sterilized transfer loop, a paper clip bent into a loop and sterilized, or a sterilized toothpick. Use the device to touch a colony of bacteria from a petri dish or test tube. Spread the bacteria on the plate in a zig-zag pattern to obtain individual colonies as the concentration of bacteria on the transfer device becomes less. Incubate the plates at 37o C for 24-36 hours. Colonies should grow to the size of this 0 for use in the lab …show more content…
Results: (For the group of this lab report)
Petri dish 1- LB and B1- Lawn growth all over the dish
Petri dish 2- LB AMP and B1-
Petri dish 3- LB and B2- Lawn growth all over the dish
Petri dish 4- LB AMP/ Mac and B2 (control) – showed no signs of grown at all
Petri Dish 1- lawn growth all over Mac B1 Petri Dish 2- Mac/ Amp B1 350 Colonies
Petri Dish 3- Lawn growth Mac B2 Petri Dish 4- Control Mac/ Amp no growth
Conclusion:
All of the groups in the class got the correct results, which means, no one had any contaminations. Some groups had more than colonies on Petri dish 2, and that just depends on how much the drops that were put on or grew. When we did the drops, not all of the drops grew because only certain plasmids took on the gene for resistance. When you have lawn growth on a plate, it means there are no obvious colonies, because they are all over on the plate. Petri dish 1 and 3 were lawn growth just as expedited. Petri dish 4 was our control and had no growth on it that the group could tell; which is what was expedited. All groups had no growth in the control plate; which once again means no