Abstract
Plasmids are circular clusters of DNA that carry valuable genes that help a host survive. The process of transformation can occur and the genetic material of the cell can be altered with a plasmid. However, the cell must be competent so it has the ability to take in DNA from the environment in order for transformation to occur properly. The E.Coli was treated with CaCl2 in order for the cell to be competent. The presence or absence of ampicillin is used to test the E.Coli’s competence for transformation. Plasmids such as lux and control pUC18 allow the cell to transform in order to grow and survive, while both contain ampicillin-resistant genes. Both the lux and control pUC18 plasmid were added to the competent bacteria in the cold ice bath.
After the ice bath, the cells were immediately transferred into a heat shock. Heat shock expands the cell allowing the plasmids to enter the host cell. The solutions were later poured into six different agar plates. Three of the agar plates contained ampicillin and the other three do not contain ampicillin. The control pUC18, lux, and competent cell solutions were split between the six agar plates. A portion of each solution was poured onto its own agar plate. Half was poured onto an agar plate with …show more content…
A cell spreader was dipped into ethanol, passed across the flame of an ethanol lamp and then cooled for about thirty seconds in order for these solutions to be poured into their respective plates. The cell spreader was used to distribute the solution evenly across the agar plates once it was cooled. For each of the plates, the process was repeated. A plate labeled “LB/Ampc” contained ampicillin and had 130 μL of the control poured onto it. Another plate labeled “LBC” had 130 μL of the mixed control poured on the agar plate from the “C” tube. The other plate labeled “LBNP” contained 130 μL of the
Plasmids are circular clusters of DNA that carry valuable genes that help a host survive. The process of transformation can occur and the genetic material of the cell can be altered with a plasmid. However, the cell must be competent so it has the ability to take in DNA from the environment in order for transformation to occur properly. The E.Coli was treated with CaCl2 in order for the cell to be competent. The presence or absence of ampicillin is used to test the E.Coli’s competence for transformation. Plasmids such as lux and control pUC18 allow the cell to transform in order to grow and survive, while both contain ampicillin-resistant genes. Both the lux and control pUC18 plasmid were added to the competent bacteria in the cold ice bath.
After the ice bath, the cells were immediately transferred into a heat shock. Heat shock expands the cell allowing the plasmids to enter the host cell. The solutions were later poured into six different agar plates. Three of the agar plates contained ampicillin and the other three do not contain ampicillin. The control pUC18, lux, and competent cell solutions were split between the six agar plates. A portion of each solution was poured onto its own agar plate. Half was poured onto an agar plate with …show more content…
A cell spreader was dipped into ethanol, passed across the flame of an ethanol lamp and then cooled for about thirty seconds in order for these solutions to be poured into their respective plates. The cell spreader was used to distribute the solution evenly across the agar plates once it was cooled. For each of the plates, the process was repeated. A plate labeled “LB/Ampc” contained ampicillin and had 130 μL of the control poured onto it. Another plate labeled “LBC” had 130 μL of the mixed control poured on the agar plate from the “C” tube. The other plate labeled “LBNP” contained 130 μL of the