1. AGENDA Who Done It Workshop Continental Breakfast Welcome and Introductions Who Done It Lab Overview of SCBC Resources on MATSC Reserving Your Kit on LARS Feedback

2. Agenda Forensics Overview Pipetting: video, steps in procedure, practice, and assessment Gel Electrophoresis Part 1: video, card sort, and prepare agarose gels (follow the SOP) Practice pipetting and loading gel wells Prepare your Samples Gel Electrophoresis Part 2: load samples and run gels Analyze results and determine identity of criminal Performance Assessment: Creating a Standard Curve to identify an unknown

3. DNA Day April 14 th 2009 DNA Day

4. National Human Genome Research Institute: DNA Day April 14 th 2009 DNA Day

18. Who Done It? Before you can put your Crime Scene Investigator hat and determine WHO committed the crime, you must first lean need to how to pipette and load dyes into a gel.

19. The Basic Laboratory Tool: Micropipette

20. Part I - An Introduction to Pipetting http://www.teachersdomain.org/resource/biot11.sci.life. gen.usingmicro/

21. How do you use a micropipette? (Prepare)

22. When using a Pipette raise the tube to eye level and remember: Number in red represents the decimal place for the P-20 Use the knob on the handle to change the volume Always use the pipette with a tip Feel the pressure difference between first stop and second stop First stop aspirates (draws up the solution) Second stop dispenses (pushes out) the solution

23. Learning a New Skill: Micropipetting (Practice)

24. Setting the Volume on a Micropipette: Think (Assessment) 1)The smallest amount of volume that I can measure with a P-20 pipette is… 2) When I use a P-20 pipette the maximum volume I can measure is…

25. Thoughts?

26. Gel Electrophoresis

27. Making an Agarose Gel http://ca.pbslearningmedia.org/resource/biot11.sci.life.gen.agarosemake/making-an- agarose-gel//

28. Electrophoresis and Gel Analysis http://ca.pbslearningmedia.org/resource/biot11.sci.life.gen.isolatedna/electrophoresis-and-gel- analysis/

29. Follow the SOP to Melt and Pour the Agarose Gel

30. Place in the microwave

31. It is ready to pour when it is cool to the touch… The solution should become clear…

32. Use the 6 well side of the Comb

33. Pour the agarose and let it solidify Remove the comb after the gel has solidified

34. Gel Electrophoresis Card Sort

35. Part II: Preparing the Samples

36. Practice Loading Dye into a Gel: Fill with 1X SB Buffer

37. Place the gel tray in the chamber with the wells closest to the negative end Pour buffer into the chamber so that it covers the gel completely

38. Load the wells, be careful not to poke a hole…

39. Place the hood on top of the chamber and press run button on the carriage Let it run for 10-15 minutes

40. Part III: Analyze the Results and Determine who Committed the Crime

41. CSS2VS1 Unknown

42. Part IV: Use mathematics to Create a Standard Curve and Identify an Unknown (Performance Assessment)