IND-27 Nucleosome Lab Worksheet (1)

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Feb 20, 2024

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Cell Biology (BIOL 215) Nucleosome Lab Worksheet Name: ___________________________ This is what our SDS-PAGE gel would have looked like if we had time to run it: We would have measured the distances that our protein standards traveled as well as each of the histones. This data is provided below. Protein Name MW Daltons Log of MW Migration on Gel (mm) Number of AA Albumin (Dimer) 132,000 7 Albumin (Monomer) 66,000 24 Ovalbumin 43,000 30 Lysozyme 14,400 56 H1 36 H5 39 H3 52 H2B 53 H2A 54 H4 57 1. Calculate the Log of the MW for the standards. (This will make it easier to graph)

2. Graph the Log MW of the standards (Y-axis) versus the migration on the gel (mm) below. Make a best fit line connecting your points. 3. Determine the Log of the MW for each unknown histone and label on your table using the migration on the gel given for each. 4. Next take the 10 X to find out the MW of each unknown in daltons and label on the table. 5. Finally, if you know that the average molecular weight for one amino acid is 120 Daltons, calculate roughly how many amino acids you would find in each protein. Plot Graph (Label appropriately):

Draw a picture of your DNA gel in the space below: a. What are the relative amounts of each protein compared to one another? b. Identify the DNA bands derived from one nucleosome versus two or three on your gel (mono, di, tri). c. Note that the mono-nucelosome bands become slightly smaller with increasing micrococcal nuclease digestion. Explain why?

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