Lab Results

only need *** DNA standard curve and DNA gel prediction: #2,3,4 answered please!!!! will upload more in 1 hr 20 mins

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The discussion section is where you interpret the results that you presented in your figures and provide conclusions for the experiments. You will use the figures that you made summarizing your data as references for your Discussion. You do not need to repeat the information in the figures and their legends, but you need to provide explanations for what the data mean. There are some experiments that you did not make a figure for that you need to explain. The discussion section should include explanations and conclusions for:
Transformation table:
1. The transformation experiment. Did the results match your predictions? What is your conclusion about the transformation? What is the difference between cells with and without pGLO? How are we able to tell which cells have pGLO and? How can we make the E. coli cells produce GFP?

***DNA standard curve and DNA gel prediction:
2. What is the relationship between the size of a DNA fragment and how far it migrates in a DNA gel? How did you use the predicted size of a DNA fragment to determine its expected migration distance on the DNA gel?
3. How many fragments did you expect to see for each of the restriction enzyme digestion reactions? What were their sizes?
4. How many fragments did you expect to see for the PCR reactions? What were their sizes?

DNA gel results:
5. Plasmid purification. Did you successfully purify the pGLO plasmid? How can you tell? How did you ensure that the culture of E. coli would maintain the pGLO plasmid for purification?
6. Restriction enzyme digestion. Were the restriction fragments on your gel the same as the fragments that you predicted? What difference(s) did you observe, if any? Explain any difference between the migration of the uncut (circular) plasmid compared to the migration of the whole, but linear plasmid (EcoR I enzyme only digestion).
7. PCR. Did you successfully amplify a portion of the pGLO plasmid? How can you tell? Was the PCR fragment the expected size? What was the result of the negative control PCR reaction?
GFP purification:
8. Did you successfully purify the GFP protein from the E. coli cells? How can you tell? How did you ensure that the culture of E. coli would maintain the pGLO plasmid and produce enough GFP to purify?