In relation to DNA Isolation experiment 1. Once the tissue has been ground and heated to 60°C, the DNA is released into the solution, but so are many other types of cellular molecules. List some types of molecules besides DNA that you would expect to find in a cell. 2. What is the role of the detergent in this protocol? How does it perform this function? 3. Name two important functions of the proteases? 4. If you wanted to isolate a copy of the gene that codes for a protein found in the skin, could that gene be located in liver cells? Explain your reasoning 5. What do you think will be the first step in purifying DNA from intact isolated cells? 6. What happened to the other macromolecules once the DNA was precipitated out of solution?

5. The proteins listed below are separated by (1) isoelectric focusing (IEF) followed by (2) sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE). A single sample of the protein mixture is placed at the top center of the gel to begin step 1. ID 1 2 3 4 5 6 7 8 Protein Collagen y-Globulin Insulin Fibronectin Lysozyme Pepsin Serum Albumin Myoglobin Molecular Weight 300,000 150,000 5,700 220,000 14,300 34,500 66,500 16,700 Conc pl 0.5 mg/L 4.7 1 mg/L 7.2 12 mg/L 5.3 5.8 11.0 5 mg/L 2 mg/L 10 mg/L - added prior to running IEF? left out of the process entirely? 4 mg/L 2 mg/L 1.0 4.9 7.1 A. Which of these proteins would electrophoresis NOT separate effectively? B. Which of these proteins would isoelectric focusing NOT separate effectively? Type fibrous a-helices globular protein globular protein fibrous protein enzyme digestive enzyme blood protein muscle protein C. Show the expected results after each step, by drawing qualitative diagrams similar to those shown in the class…

DNA EXTRACTION Why do scientists isolate DNA?                                                                                                                    From what part of the subject’s body were cells collected? Give the specific purpose of each experimental step:           Lysis solution & heat:                                                                                             Salt & centrifugation:                                                                                  After centrifuging, where in the tube is the DNA? What is the purpose of adding alcohol?