Data
LB-Plasmid
Prediction: No the plate will not show growth because there is no foreign DNA.
Observation: There was a large amount of bacteria growth.
Observation: There was a large amount of bacteria growth.
LB+Plasmid
Prediction: Yes the plate will show grow because it has foreign DNA
Observation: There was also a large amount of bacteria growth.
Observation: There was also a large amount of bacteria growth.
LB/Amp-Plasmid
Prediction: No the plate will not show growth because there is no foreign DNA.
Observation: There was no bacteria growth.
Observation: There was no bacteria growth.
LB/Amp+Plasmid
Prediction: Yes the plate will show growth because it has foreign DNA.
Observation: There was a small amount of bacteria growth.
Observation: There was a small amount of bacteria growth.
Analysis
4.) The number of individual colonies in each.
LB+plasmid (positive control) = lawn
LB-plasmid (positive control) = lawn
LB/Amp+plasmid (experiential) = 19 colonies
LB/Amp-plasmid (negative control) = none
LB-plasmid (positive control) = lawn
LB/Amp+plasmid (experiential) = 19 colonies
LB/Amp-plasmid (negative control) = none
5.) Compare and contrast number of colonies on each.
LB+plasmid and LB-plasmid - The recombinant DNA did not affect either.
LB/Amp-plasmid and LB-plasmid - The ampiciline kills most of the bacteria.
LB/Amp+plasmid and LB/Amp-plasmid - The recombinant DNA prevents the plasmids from dying.
LB/Amp+plasmid and LB+plasmid - The ampaciline still restricts growth but does not prevent it.
LB/Amp-plasmid and LB-plasmid - The ampiciline kills most of the bacteria.
LB/Amp+plasmid and LB/Amp-plasmid - The recombinant DNA prevents the plasmids from dying.
LB/Amp+plasmid and LB+plasmid - The ampaciline still restricts growth but does not prevent it.
6.) For this experiment, we are selecting the antibiotic resistance.
7.) N/A. The phenotype did not work.
8.) I would first inspect the LB/Amp+ plate to conclude that the transformation occured successfully because the bacteria grows in the presence of the antibiotic.
9.) Transformation efficiency
A.) The total mass of plasmid used is .05 ug.
B.) The total volume of cell suspension prepared is 510 uL.
C.) The fraction of the total cell suspension that was spread on the plate is 10/51.
D.) The mass of plasmid in the cell suspension spread is .0098.
E.) The number of colonies per ug plasmid DNA is .1863.
B.) The total volume of cell suspension prepared is 510 uL.
C.) The fraction of the total cell suspension that was spread on the plate is 10/51.
D.) The mass of plasmid in the cell suspension spread is .0098.
E.) The number of colonies per ug plasmid DNA is .1863.
10.) Influencing factors and their effects
The factors that influence the transformation efficiency could have possible been the beads. The beads may not have been rolled over enough thus not sufficiently spreading the plasmid to the e. coli.