BIO 201 Research Module #1 - Week 3.pdf - Week 3 Protein Content Analysis and Study Design Last week you used a number of analytical procedures to

BIO 201 Research Module #1 - Week 3.pdf - Week 3 Protein...

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Week 3 - Protein Content Analysis and Study Design Last week, you used a number of analytical procedures to gather data to help you work toward determining the identities of a series of five unknowns. You now havesome evidence that has probably allowed you to make informed deductions about the identity of at least some of the unknowns. However, you likely still need more data to be able to make a strong evidence-based argument about the identity of all the unknowns. Quickly complete background research to determine levels of protein expected in each of the following solutions - grape juice, kosher wine, non-kosher wine, blood and Kool Aid. Accepting this piece of information will potentially permit you to discriminate between the unknowns, specifically between kosher wine, non-kosher wine, and Kool Aid. In this procedure, you will use spectrophotometry to determine protein concentrations your unknowns. This will require creating a standard curve of absorbance as a function of protein level, then using that relationship to determine protein content in the 5 unknown samples. (This is similar to last week’s exercise with xylene cyanol.) Analyze protein content in the unknowns by performing a protein assay. You will construct a standard curve of known protein concentrations (mg/mL) on the X-axis versus absorbance at 595nm on the Y- axis. You will use this standard curve to quantify the protein concentration in each of the 5 unknowns. The protein assay that you will use is a coomassie blue binding assay. Coomassie blue is a dye that binds to one particular amino acid; recall that amino acids are the units that link together to form proteins. When coomassie blue binds to the amino acid (or more generally, the protein), the dye changes color from brown to blue. The change in color reflects a change in spectral/absorbance properties of bound coomassie blue
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