Kim Zwier

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Kim Zwier

• Dr. Macdonald Wicks Lab
• Muscle growth and development

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My Research Project

• Identify muscle proteins associated with growth
  in yellow perch

Perca flavescens
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From Fish to Protein Separation
Muscle Sample
Protein Separation
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Why this Research is Important

• If we can understand protein patterns in yellow
  perch, then we can understand it better in other
  animals

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Why Yellow Perch?

• Yellow perch is now being selected for the
  quantitative trait of growth

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Experimental Design

• 132 samples evaluated from 8 age groups
• 9,10, 11, 12, 16, 20, 26, and 28 weeks
• Body weight and length
• All samples taken from same strain across ponds
  to eliminate confounding variables
• Water content
• What fish eat

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Yellow Perch Muscle Fingerprint in Different
Concentrations

• Different dilutions were tested
• 13. 19, 127, and 181
• This allows us to detect the full range of
  proteins

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My Daily Activities

• Prepare and run the gel
• Record data
• Scan gels using alpha innotech scanner
• Analyze the gels on Phoretix software

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Preparing and Running the Gel

• Prepare the 5 and 20 gradient gels
• Put the gel plates together
• Pour the gel through the gradient maker
• Make the stacking gel
• Load gel with centrifuged samples
• Run the gel on 500 volts for about 2.5 hours

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Putting the Gel Plates Together
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Preparing the Gel
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The Gel Plates with Comb
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The Buffer Tanks
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Electrophoresis

• Separates proteins by size using an electric
  field
• Largest to smallest

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Coomassie Blue Stain

• Blue dye was added to the 20 gradient gel and
  5 stayed clear

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Scanned Gel (1D Fingerprint)
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Analyzing the Gel Fingerprint
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From Fish to Protein Separation
Muscle Sample
Protein Separation
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Results and Conclusions

• More proteins can be found with different
  concentrations.
• We have found that there is a direct linear
  correlation with increase in the concentration of
  certain muscle proteins and the size of yellow
  perch.

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What I Learned

• How to work safely in a laboratory environment
• How to make a gel
• How to do electrophoresis
• How to use the Phoretix computer program
• Efficient planning
• Analytical reasoning

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Questions?