E. Coli Transformation with Plasmid (Pgal), Pgal Isolation, and Analysis of Plasmid Dna
E. Coli Transformation with Plasmid (pGal), pGal Isolation, and Analysis of Plasmid DNA
Felicia Osadi
Bio 22
April 20, 2012
Transformation = group 10
Plasmid = group 7
RFLP = group 1
RESULTS
Table I. Plasmid Transformation of E. Coli. Plate # | Agar plate | Type | Result | 1 | X-gal | Control | Extensive lawn growth | 2 | Ampr / X-gal | Control | Clear no bacterial growth | 3 | Ampr / X-gal | Transformation | 1 blue colony |
Transformation efficiency = 1 transformants x 1.0 mL = 200 transformants/ug 0.025 ug 0.2 mL
Figure 1: Plasmid Isolation Gel Electrophoresis. This figure represents result from several groups doing plasmid isolation. All the data are collected in Bio 22 class on February 24, 2012. DNA obtained from this experiment comes from a plasmid DNA that were isolated and put into a buffer solution before running it through a gel. The result of our particular experiment is in lane 2. The sample in lane 2 is a control group which contains RNAse instead of DI water. This particular gel is 0.8% agarose.
Figure 2: Restriction Fragment Length Polymorphism of Circular DNA. Experiment which resulting this figure is done in Bio 22 lab on April 6, 2012. This figure represents several data that are obtained from adding certain restriction enzyme. Lane #1 represents a ladder, a marker of known fragment of DNA. Lane #2 represents uncut circular DNA. Lane #3 and #4 is the same circular DNA treated with SmaI and XhoI respectively. Solution in line #5 is treated with both SmaI and XhoI. While adding enzyme, the total volume is kept constant at 50 µL by adding water to the solution.
Figure 3: Standard Curve of Control Group in RFLP. All the data in this curve are obtained from the ladder that is used in RFLP experiment. Distance migrated is obtained by measuring the distance in cm from the top of the gel to the point where it migrated. The numbers of base pairs are given to us for
Felicia Osadi
Bio 22
April 20, 2012
Transformation = group 10
Plasmid = group 7
RFLP = group 1
RESULTS
Table I. Plasmid Transformation of E. Coli. Plate # | Agar plate | Type | Result | 1 | X-gal | Control | Extensive lawn growth | 2 | Ampr / X-gal | Control | Clear no bacterial growth | 3 | Ampr / X-gal | Transformation | 1 blue colony |
Transformation efficiency = 1 transformants x 1.0 mL = 200 transformants/ug 0.025 ug 0.2 mL
Figure 1: Plasmid Isolation Gel Electrophoresis. This figure represents result from several groups doing plasmid isolation. All the data are collected in Bio 22 class on February 24, 2012. DNA obtained from this experiment comes from a plasmid DNA that were isolated and put into a buffer solution before running it through a gel. The result of our particular experiment is in lane 2. The sample in lane 2 is a control group which contains RNAse instead of DI water. This particular gel is 0.8% agarose.
Figure 2: Restriction Fragment Length Polymorphism of Circular DNA. Experiment which resulting this figure is done in Bio 22 lab on April 6, 2012. This figure represents several data that are obtained from adding certain restriction enzyme. Lane #1 represents a ladder, a marker of known fragment of DNA. Lane #2 represents uncut circular DNA. Lane #3 and #4 is the same circular DNA treated with SmaI and XhoI respectively. Solution in line #5 is treated with both SmaI and XhoI. While adding enzyme, the total volume is kept constant at 50 µL by adding water to the solution.
Figure 3: Standard Curve of Control Group in RFLP. All the data in this curve are obtained from the ladder that is used in RFLP experiment. Distance migrated is obtained by measuring the distance in cm from the top of the gel to the point where it migrated. The numbers of base pairs are given to us for