Plasmid DNA Transformation Lab Report

Due to limited time constraints and having lab only once a week this lab was broken up into five periods, the last one specifically to go over the results. Day one consisted of extraction of plasmid DNA, which we used Resuspension Buffer to break up our pellet after removing the supernatant (liquid) after centrifuging our overnight culture. Then Lysis Buffer was used to break down the membrane of the cells so that we could then add Neutralization Buffer to bring our sample from basic to neutral pH. After centrifuging a few times with some steps in between we added Wash Buffer to remove all other liquid from the spin column leaving just the DNA in its membrane. Later some Elution Buffer was used to separate the DNA from the membrane of the spin column. Day two consisted of making the plasmid DNA resistant to Kanamycin and Ampicillin. Day three we made 0.8% agarose gel and ran electrophoresis to be able to see the weight of the DNA. Day four we genetically transferred the ligated plasmid DNA to E.coli and plated the cell mixture. Day five we looked at our results and went over them. All of this works together for bacterial transformation, we transformed E.coli to be Kanamycin and Ampicillin resistant, by adding the Kanamycin and Ampicillin plasmid DNA to E.coli.
Procedure:
Start this lab by isolating plasmid DNA from E.coli so that we can use it in …show more content…
First we ran a gel electrophoresis to make sure that we had obtained the DNA like we thought. Once the electrophoresis was done we could clearly see that we did have DNA (Figure 1). Then we added the DNA to the ligated E.coli, once this was done we added the samples to a plates that had Kanamycin and Ampicillin in the agarose to see if we were at all successful in making the E.coli also resistant to Kanamycin and Ampicillin. Looking the plates we can see many colonies grew, so we were again successful (figure