Ligation Test Lab Report
The Effect of Varying Ligation Conditions on Ligation Efficiency
Introduction The goal of this experiment was to successfully ligate RFP into P1 and transform E. coli with P1-RFP plasmid, which contains the P1 promoter, RBS, and RFP gene. We tested several different ligation conditions and assessed how they each affected ligation efficiency. The two variables that we tested were: molar ratio of insert (RFP): vector (P1 plasmid), as well as the effect of removing the stuffer from the digested plasmid. We predicted that the cells transformed without stuffer would have higher ligation efficiency than the cells transformed with stuffer. The reason for this is because the stuffer (if not removed) would cause the plasmid to ligate back into itself …show more content…
Average ligation efficiencies under varied ligation conditions. The average ligation efficiencies (conditions listed from left to right in the figure) were: 39.81%, 42.80%, 58.98%, and 62.72%, respectively. By removing the stuffer from the plasmid showed an increase in ligation efficiency for both 1:1 and 2:1 insert: vector conditions. Having 2:1 insert: vector also showed an increase in ligation efficiency for both with stuffer and without stuffer.
Table 1. Standard deviations of each ligation data set. This illustrates the variation in ligation data collected from individual groups. Compared to the average ligation efficiencies (refer to error bars on Fig. 1), the data of individual groups for each ligation condition (listed from left to right in the table) ranged from: 19.6-59.2%, 11.8-76.0%, 25.0-92.3%, and 12.5-89.8%, respectively. From this, we can generally see how the data from each group is spread out across each ligation condition.
Table 2. ANOVA test results. The p-value corresponding to the F-statistic is lower than 0.05, suggesting that one or more treatments are significantly different. We can see that the treatments that are significantly different are the 1:1 with stuffer and 2:1 without stuffer.
Introduction The goal of this experiment was to successfully ligate RFP into P1 and transform E. coli with P1-RFP plasmid, which contains the P1 promoter, RBS, and RFP gene. We tested several different ligation conditions and assessed how they each affected ligation efficiency. The two variables that we tested were: molar ratio of insert (RFP): vector (P1 plasmid), as well as the effect of removing the stuffer from the digested plasmid. We predicted that the cells transformed without stuffer would have higher ligation efficiency than the cells transformed with stuffer. The reason for this is because the stuffer (if not removed) would cause the plasmid to ligate back into itself …show more content…
Average ligation efficiencies under varied ligation conditions. The average ligation efficiencies (conditions listed from left to right in the figure) were: 39.81%, 42.80%, 58.98%, and 62.72%, respectively. By removing the stuffer from the plasmid showed an increase in ligation efficiency for both 1:1 and 2:1 insert: vector conditions. Having 2:1 insert: vector also showed an increase in ligation efficiency for both with stuffer and without stuffer.
Table 1. Standard deviations of each ligation data set. This illustrates the variation in ligation data collected from individual groups. Compared to the average ligation efficiencies (refer to error bars on Fig. 1), the data of individual groups for each ligation condition (listed from left to right in the table) ranged from: 19.6-59.2%, 11.8-76.0%, 25.0-92.3%, and 12.5-89.8%, respectively. From this, we can generally see how the data from each group is spread out across each ligation condition.
Table 2. ANOVA test results. The p-value corresponding to the F-statistic is lower than 0.05, suggesting that one or more treatments are significantly different. We can see that the treatments that are significantly different are the 1:1 with stuffer and 2:1 without stuffer.