Name: _____________________________________________Date: __________________________ Period: ___________ LAB: CSI Halloween Macromolecules Crime Scenario Background: Mrs. Nichol’s Mad Scientist twin sister, Dr. Barb Cooper, has been secretly growing genetically engineered pumpkins in Mrs. Nichol’s stockroom at Curwensville, without Mrs. Nichol’s knowledge. Dr. Barb added a special firefly gene to the DNA of the genetically engineered pumpkins. The gene, called luciferase (meaning light), produces an enzyme (protein) which makes the pumpkins glow. Dr. Barb will be a millionaire if she can market the pumpkins for Halloween. Dr. Barb, unlike Mrs. Nichol, was not very smart. She began bragging about her pumpkin discovery to her 2 questionable friends: Christy “Wheat Bread” Jones, occupation: Bread Baker and Ted “T-Bone Steak” Smith, occupation: Meat Butcher. Using your knowledge of macromolecules (Carbohydrates, Lipids and Proteins), you will need to determine who the thief or thieves were based on the crime scene evidence. Crime Scene Preliminary Evidence Police Report: This morning, 10/29/2010, at 7:00am, Mrs. Nichol entered her classroom to find the stockroom door open and a smashed genetically engineered pumpkin on her floor, oozing an orange substance. Wet orange footprints were found near the scene. A smudgy substance was found on the door handle to the stockroom. No fingerprints or DNA were found. Investigation: As a CSI (Crime Scene Investigator) specializing in macromolecules, you will need to analyze the evidence found at the crime scene to determine which suspect may have stolen the genetically engineered pumpkin from Mrs. Nichol’s stockroom. Use the lab data collected and the information in the suspect report to help you. It may be only one or both suspects who are guilty. When you have decided on a suspect, you will need to turn in a warrant for their arrest (attached to the lab sheets). Mrs. Nichol, who moonlights as a judge, will approve or deny the warrant based on the strength of evidence you put together in your investigation. Make sure you don’t wrongly accuse someone who is innocent!!! Suspect Report: The suspects are Dr. Barb’s two questionable friends: 1. Christy (AKA “Wheat Bread”) Jones, occupation: Bread Baker Always covered in flour and starch from baking Orange substance found on shoes at house – claims she stepped in orange Jell-O split on her kitchen floor 2. Ted (AKA “T-Bone Steak”) Smith, occupation: Meat Butcher Clothes and shoes usually contaminated with meat and fat Orange substance found on shoes at house – claims he stepped in orange juice spilt on his kitchen floor Evidence List: Your evidence list will contain the following: 1. Sample of orange ooze from smash pumpkin at crime scene 2. Sample of smudgy substance found on stockroom door handle 3. Samples of orange substances found both suspects shoes a. Christy (AKA “Wheat Bread”) Jones, occupation: Bread Baker b. Ted (AKA “T-Bone Steak”) Smith, occupation: Meat Butcher Safety: 1. Wear goggles at all times. 2. Wear an apron at all times. 3. Solutions, stains, and reagents are dangerous. They may irritate the skin or cause staining of the skin and/or clothing. 4. Use extreme care when working with heated equipment or materials to avoid burns. 5. NEVER handle hot glassware without proper equipment. 6. Be careful to avoid breakage when working with glassware. 7. Wash hands with soap and water when you are finished with the lab. 8. IF IN DOUBT...ASK Mrs. Nichol!!! 1 Name: _____________________________________________Date: __________________________ Period: ___________ Outline: 1. You will need to test the crime scene evidence to determine the macromolecules present (carbs, lipids and proteins), in the unknown substances at the crime scene: (the orange substance oozing from the pumpkin and the smudgy substance on the door handle). 2. On your CHART, Make a hypothesis of what each substance may be based on the macromolecule test results and the suspects’ occupations (starch, meat, fat, etc). 3. You will then test each suspect’s shoes to see if the orange substance on the shoes matches that found at the crime scene oozing from the genetically engineered pumpkin. 4. You will submit a warrant for the suspect(s) you think is/are guilty to Judge Nichol with evidence to support your claim. She will approve or deny the warrant based on your evidence. Pre-Lab Questions: Read the lab sheets & answer the questions using the lab and your Organic Compound Notes/ Chart. 1. Explain all safety precautions necessary in lab. __________________________________________________________________________________________ __________________________________________________________________________________________ __________________________________________________________________________________________ 2. What are the four types of macromolecules/organic molecules? __________________________________________________________________________________________ 3. Name the three types of carbohydrates. What type(s) of carbohydrate is tested for in this lab? How do you test for this type of carbohydrate? _______________________________________________________________________________ __________________________________________________________________________________________ __________________________________________________________________________________________ __________________________________________________________________________________________ 4. Name two types of proteins found in the “Crime Scene Scenario Background”. How do you test for proteins? __________________________________________________________________________________________ __________________________________________________________________________________________ __________________________________________________________________________________________ 5. Name 3 types of lipids. How do you test for lipids? __________________________________________________________________________________________ __________________________________________________________________________________________ __________________________________________________________________________________________ 6. What macromolecule(s) would Ted (T-bone steak) Smith leaves behind at a crime scene? __________________________________________________________________________________________ 7. What macromolecule(s) would Christy (Wheat Bread) Jones leave behind at a crime scene? __________________________________________________________________________________________ 8. What macromolecules might be contained in the genetically engineered pumpkin? Why? __________________________________________________________________________________________ _____________________________________________________________________________________ 2 Name: _____________________________________________Date: __________________________ Period: ___________ Procedure: You will need to test each substance for carbohydrates, lipids and proteins. ***You will be testing each of the 4 evidence substances for Simple & Complex Carbohydrates (8 test tubes), Lipids (4 test tubes), & Proteins (4 test tubes) = 12 test tubes. Set this up the day before the lab: 1. Take a Sharpie and label each one of your test tubes the following: 1SC/ 2SC/ 3SC/ 4SC Simple Carbohydrates for each piece of evidence. 1CC/ 2CC/ 3CC/ 4CC Complex Carbohydrates for each piece of evidence. 1L/ 2L/ 3L/ 4L Lipid Test for each piece of evidence. 1P/ 2P/ 3P/ 4P Protein Test for each piece of evidence 2. Get 8 little beakers (100 mL). a. Label four as: 1; 2; 3; 4-when you do the lab the 4 orange evidence substances will go in these-see chart on page 5 for what each number represents. When you perform the lab stir then pour a small amount from the stock beaker of each orange substance in the appropriate labeled beaker to use at your table. Put a clean pipette in each beaker. b. Label the other four 100 mL beakers as: Iodine; Benedict’s; Sudan III; Biuret’s-when you perform the lab put a small amount of the reagent from the stock bottle into the appropriate labeled beaker to use at your table & have a new pipette for each beaker. 3. Use a Sharpie & ruler to set up bag for lipid test- see lipid procedure #7 Lab tests Procedures: Complex (Polysaccharide) Carbohydrate test: 1. Obtain test tubes 1CC – 4CC & add 5mL (10 drops) of the each substance to its correct test tube. 2. Place each test tube back into the test tube rack. 3. Add 5 drops of Iodine solution to the first test tube (1CC). Caution: Iodine solution is poisonous and will stain your hands and clothes if you aren’t careful. 4. Carefully swirl contents making sure not to spill any of the solution. 5. Record a positive (+) or negative (-) result for the Iodine solution test, on your data table. If the solution changes from YELLOW-BROWN to BLACK-PURPLE, starch is PRESENT. 6. Repeat steps 3-5 with test tubes 2CC-4CC. 7. Make sure to take a video of test tube reactions, then dump & rinse test tubes with hot water down an EMPTY SINK and wash test tubes immediately in soapy water and rinse well. Run hot water down drain several minutes. Simple (Monosaccharide) Carbohydrate Test 1. 2. 3. 4. Obtain test tubes 1SC – 4SC & also label with your group’s name. Add 5mL (10 drops) of the each substance to its correct test tube. Place each test tube back into the test tube rack. Add 5 drops of Benedict’s solution to each test tube. a. Carefully swirl contents making sure not to spill any of the solution and place tubes in hot water bath 10 minutes Caution: Water is hot –do not get burned! MOVE ON TO THE LIPID TEST NOW!!! b. If the substances turned green, yellow, orange or red after 5 minutes, then a monosaccharide is present. Record a + or - for the Benedict’s test on the data table for substance. NOTE: Since your substance is orange and one of the indicators of a positive test result is orange, you may need to compare your finished sample to the original sample to see if there is a significant color change. c. Make sure to take a video of test tube reactions, then dump & rinse test tubes with hot water down an EMPTY SINK and wash test tubes immediately in soapy water and rinse well. Run hot water down drain several minutes. 3 Name: _____________________________________________Date: __________________________ Period: ___________ Lipid test: 1. Obtain test tubes 1L – 4L & add 5mL (10 drops) of the each substance to its correct test tube. 2. Place each test tube back into the test tube rack. 3. Add 5 drops of Sudan III stain to the first test tube (1L). Caution: Sudan III stain will stain your hands and clothes if you aren’t careful 4. Carefully swirl contents making sure not to spill any of the solution. 5. Record a positive (+) or negative (-) result for the Sudan III stain test, on your data table. If the solution changes to RED, then a LIPID is present. Repeat steps 3-5 with test tubes 2L-4L. 6. Make sure to take a video of test tube reactions, then dump & rinse test tubes with hot water down an EMPTY SINK and wash test tubes immediately in soapy water and rinse well. Run hot water down drain several minutes. 7. Take a brown paper bag & divide the bag into 4 parts using a ruler and Sharpie. 8. Label each part according to your evidence Pumpkin/ Door-knob/ Bread Baker/ Meat Butcher & put your names on the bottom of the bag. 9. using a different Popsicle stick for each substance, rub a small portion of each substance onto the brown paper bag. Wait for 2 minutes…. 10. Record a positive (+) or negative (-) result on the data table for each substance. If the section of the bag is TRANSLUCENT, a lipid is PRESENT. 11. Make sure to take a video of bag results. Protein test: 1. Obtain test tubes 1P – 4P & add 10drops each substance to its correct test tube. 2. Place each test tube back into the test tube rack. 3. Add 10 drops of Biuret’s reagent to the first test tube (1P). Caution: Biuret’s solution contains a strong base. If you get any of this solution on your skin, wash with soap and water immediately. Have your partner get me immediately as well 4. Carefully swirl contents making sure not to spill any of the solution 5. Record a positive (+) or negative (-) result for the Biuret’s reagent test, on your data table. If the solution changes from YELLOW to BLUE-VIOLET, then the protein is PRESENT. 6. Repeat steps 3-5 for tubes 2P-4P. 7. Make sure to take a video of test tube reactions, then dump & rinse test tubes with hot water down an EMPTY SINK and wash test tubes immediately in soapy water and rinse well. Run hot water down drain several minutes. 4 Name: _____________________________________________Date: __________________________ Period: ___________ RESULTS CSI Macromolecule Data Table: Test Substance Predictions (does the evidence include simple carbs/complex carbs Lipids/proteins) Complex Carbohydrate +/- Simple Carbohydrate +/- Lipids Sudan III +/- Lipids (brown Protein +/- bag) +/- Pumpkin (1) Doorknob (2) Bread Baker (3) Meat Butcher (4) Additional Notes-Observations: 5 Name: _____________________________________________Date: __________________________ Period: ___________ OFFICIAL WARRANT CHS CSI Division We, the CSI Macromolecule specialist team of CHS, hereby believe suspect(s) ___________________________________________________________________ Are guilty of the crime ________________________________________________ ___________________________________________________________________ Based on the evidence that _____________________________________________ ___________________________________________________________________ ___________________________________________________________________ ___________________________________________________________________ ___________________________________________________________________ ___________________________________________________________________ ___________________________________________________________________ ___________________________________________________________________ CSI Team Signatures (PRINTED) Approved _____________ ___________________________ ___________________________ ___________________________ ___________________________ ___________________________ Denied________________ Signature___________________ Judge Nichol 6