Connecting the last few labs

The last several labs have been connected and here's a little summary to remind you how they are interconnected:

• Labs 7-8: Bacterial genetic transformation with pGLO plasmid
• Labs 9-10: Protein (GFP) purification by chromatography
• Labs 11-12: Bradford protein (GFP) determinations
• Lab 13: Native Protein PAGE
• Lab 14: Denaturing Protein PAGE

When we transformed our E. coli with the pGLO plasmid, containing the gene for the Green Fluorescent Protein (GFP) we were also producing the material we were going to use in the next several labs. We transformed the bacteria, made them produce our GFP, then purified that GFP and now we are determining how much of it there is and how big the polypetide is.
This process is analogous to that in the industry to produce a protein for commercial applications asn also abalogous to a research process when a gene for a new protein is being discovered or studied.

Dont think of these few labs as independent ones, but as steps of a single longer lab.

Labs 11-14 can be summarized in a single lab report. Here are the questions for such report:

1. Why do we make our absorbance measurements at 595 nm?
2. When you heat up DNA or protein samples up to 95ºC they get denatured. Specifically what happens to DNA and proteins? In other words what is DNA denaturation and what is protein denaturation?
3. When estimating protein size would you rely more on the native of the denaturing gel results? Why?
4. Some groups had more than one protein (more than just GFP) in their gels. HOw do you explain this?
   

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