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A simulation of a genetic test using Gel Electrophoresis

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1 A simulation of a genetic test using Gel Electrophoresis
Testing for Familial Hypercholesterolemia Caused by a Mutation in the LDLR Gene A simulation of a genetic test using Gel Electrophoresis

2 Nature vs. Nurture Nature = genes and their effects
Nurture = environmental influences Examples Sickle Cell Disease is heavily nature because it is related to a specific gene that are inherited Hypercholesterolemia is a blend of Nature and nurture Nature = genes such as APOB, PCSK9, LDLRAP1 and LDLR Nurture = diet, exercise Most traits are a combination of Nature and Nurture Most traits are controlled by many genes Most traits are effected my several environmental influencers

3 Human Genome We have two in every cell! Human Genome
Maternal and Paternal sources Human Genome Complete set of human DNA About 3 billion base pairs Segmented into 23 different sections called chromosomes 23(egg) + 23(sperm) = 46(zygote) We have two of every gene. Genes come in various forms. Ex: ‘wild type’ hemoglobin and sickle cell hemoglobin

4 Hypercholesterolemia
High cholesterol High cholesterol leads to Coronary Artery Disease (CAD) which often results in heart attacks.

5 Our Lab This lab tests one of the genes (LDLR) associated with high cholesterol Positive result means an 85% risk of heart attack for men in there 40s and 50s or women in 50s or 60s Negative result means average risk of heart attack Due to environmental factors (diet, exercise) Background: Parents of Sylvia (2 yr old) both have high cholesterol. They want to know if it is related to their genes and if they passed the genes to their daughter. DNA samples have been collected from Mom, Dad and Sylvia The LDLR gene in each of the samples have been amplified and digested with a restriction enzyme. We will perform the gel electrophoresis procedure, analyze the resulting gels and inform the parents of the outcome of the test and their daughters risk level of developing high cholesterol.

6 Day 1 How to use a micropipette
Look at the number on the top of your micropipette Look at the numbers on the side (through the window) Push down the top “plunger” - feel first and second stops First stop is for picking up Second stop is for dispensing Watch how to pick up a tip

7 Day 1 How to use a micropipette
Watch how to pick up a liquid Always hold the liquid in one hand while picking it up ONLY go to the first stop when picking up a liquid Watch how to dispense a liquid Always go to the first stop, then slowly to the second stop Pull micropipette out BEFORE releasing the plunger Watch the technique

8

9 Day 1 - Practice Loading ALWAYS use a TIP with your micropipette
ALWAYS keep the micropipette VERTICAL when you have liquid 1 l (micro) = 10-3 mL (milli) = 10-6 L When picking up liquid, push to the FIRST stop only! When dispensing liquid, push to the FIRST stop, then the SECOND stop When dispending liquid, ALWAYS raise the tip out of the liquid before letting go of the button!

10 Day 2 - Loading/Running Gels
Only two group members at the gel at any given point Each group member should load at least one sample Load 35 mL sample into the wells (label the wells on the diagram so you know where every sample is!) Tap the sample so that all is at the bottom of the container Let me know once all samples are loaded Work on the ‘paper lab’ activaty while you wait

11 Day 2 DNA samples Sample A – A known DNA sample called a “Standard” that contains fragments of known length Sample B – Control DNA that is known to have two normal copies of the LDLR gene Sample C – Control DNA that is known to have two mutant copies of the LDLR gene Sample D – Sylvia’s mom’s DNA Sample E – Sylvia’s DNA Sample F – Sylvia’s dad’s DNA

12 Groups Group 1 Group 2 Group 3 Group 4 Group 5 Group 6 Olivia Shanice
Oliver Emma Liam Patrick Suzanna Ashley Clara Alana Jessica Marina Ethan Mei Yarden Charles Jack Becca Mike

13 Day 3: Gel Analysis Place your gel on the light box
Do a detailed sketch of your Gel and the reference gel in your packet Include in the sketch # and position of bands Band brightness Label the lanes Use both gels when drawing conclusions about Sylvia. Only one group at the light box at one time.

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