AGENDAWho Done It Workshop

•Continental Breakfast•Welcome and Introductions•Who Done It Lab•Overview of SCBC•Resources on MATSC

• Reserving Your Kit on LARS

•Feedback

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

DNA Day April 14th 2009 DNA Day

National Human Genome Research Institute: DNA Day April 14th

2009 DNA Day

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.

The Basic Laboratory Tool: Micropipette

Part I - An Introduction to Pipetting

http://www.teachersdomain.org/resource/biot11.sci.life.gen.usingmicro/

How do you use a micropipette? (Prepare)

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

Learning a New Skill: Micropipetting (Practice)

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…

Thoughts?

Gel Electrophoresis

Making an Agarose Gel

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

Electrophoresis and Gel Analysis

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

Follow the SOP to Melt and Pour the Agarose Gel

CAUTION!!!! THE SOLUTION WILL BE EXTREMELY HOT AFTER MELTING- please use

provided mitt

1. Obtain one bottle of agarose solution as directed by your teacher.

2. LOOSEN the CAP and place in microwave for approximately 2 minutes or until agarose gel

solution is completely melted and clear.

3. Use mitt to remove melted solution from the microwave and place on the lab bench.

4. Remove the cap from the bottle and let the solution cool down to ~ 50 degrees Celsius

Place in the microwave

It is ready to pour when it is cool to the touch…

The solution should become clear…

Use the 6 well side of the Comb

Pour the agarose and let it solidify

Remove the comb after the gel has solidified

Gel Electrophoresis Card Sort

Part II: Preparing the Samples

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

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

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

Place the hood on top of the chamber and press run button on the carriage

Let it run for 10-15 minutes

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

CS S2V S1

Unknown

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