Quiz 1

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Quiz 1

• Grab a quiz sheet
• Write down section
• Take everything off desk
• You have 10 minutes

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1. What organelle functions in energy production
in animal cells. What about plants cells? 2.
Which of the following is membrane
bound? Nucleus or Nucleoid
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• 3. Which are the two modes of reproduction in
  Ciliates, such as Paramecium?
• _________________
• _________________

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4. Trypanosoma belongs to the Phylum
_________________
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5. This organism belongs to the Kingdom
_________________ and moves by means of
____________
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• 6. What is a unique feature of a parfocal
  microscope?
• a) light is formed on the slide
• b) can use both eyes to look at an image
• c) focus is retained as you move between
  objective lens
• d) one eye can be focused independently of the
  other

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7. How many structures of a protein are there?
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Spectrophotometry

• . Use of spectrophotometers.
• . Zero the machine.
• . Composition and use of a blank.
• . How to adjust wavelength.
• . Proper use of cuvettes.

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ENZYME KINETICS
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Proteins

• A protein's function is due entirely to its
  overall shape conformation.
• Conformation is determined by 1, 2, 3, and
  perhaps 4 structure of the protein.

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The primary structure of a proteinsequence of
amino acids
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The secondary structure of a proteinHydrogen
bonding between C N
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The tertiary structure of a proteinInteraction
of amino acids and H2O
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The quaternary structure of proteins2 or more
protein molecules joined together
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Enzymes

• Enzymes are protein catalysts.
• Lower the activation energy necessary for a
  chemical reaction to occur.
• Act on a substrate, form an enzyme-substrate
  complex. Ultimately results in a product.
• We are using barley amylase enzyme on a starch
  substrate.

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Formation of an enzyme-substrate complex
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Environmental factors affecting enzyme activity
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Blanking spectrophotometer

• Set wavelength to 560 nm
• With chamber empty, set Transmittance T0 with
  left knob
• Place blank cuvette in spectrophotometer
• Set Absorbance A0 with right knob
• Replace blank for experimental cuvette
• Take reading!!!

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BASIC PROCEDURE
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• 1. Prepare 2 erlenmeyers (one for pH and one for
  temperature)
• 2. Set wavelength to 560 nm
• 3. Prepare a blank and blank your spec
• 4. Collect your initial reading (Time zero)
• 5. TA will put 1 ml of enzyme in your
  erlenmeyer.
• 6. Start timing

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• 7. As the proper time interval approaches,
  swirl the flask to mix and draw 5 ml of
  starch/enzyme up into the pipette. You should
  pipette up the solution about 15 sec before the
  reading time.
• 8. At the reading time, release the solution
  into the cuvette and mix - the iodine stops the
  reaction immediately.
• 9. Readings should generally be taken
  within about 5 minutes.

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Experiments

• You will work in groups of three.
• Each group will perform one run from the pH
  experiment (4.0, 4.5, 5.0, 5.5, 6.0 and 6.5)
• Then, each group will do one of the temperature
  variables (15º, 30º, 45º, 55º, 60º and 70º)

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pH experiment

• -Add 35 ml of buffer to 35 ml of starch solution.
• -Blank is 11 dH2Obuffer ( iodine)

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Temperature experiment

• -Mix as in basic procedure. The reaction flask is
  kept in a water bath.
• -NOTE Very cold and very hot solutions might
  need to sit before being read.

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I will come around and add enzyme when your group
has

• Finished basic procedure
• 35 mL pH buffer 35 mL starch
• Blank- 0.1 mL I 2.5 mL pH 2.5 mL DI H2O
• Placed 0.1 mL I in each cuvette
• Blanked Spectrophotometer
• Taken time 0
• Transfer 5 mL solution from flask into cuvette w/
  I. Make sure to mix!

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All of the data will be shared among the class

• You will record your data on the computer.
• A data set will be printed out for each student.

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Reaction Rate

• 1/2 change in absorbance /
• time for this change to occur
• -There is a worksheet at the end of the chapter
  to help calculate reaction rate.
• Additional copies of this worksheet can be
  printed off of the homepage.
• Hand this in with your lab report, so that I can
  see your calculations.
• There is another handout ("How to Calculate
  Reaction Rates") that is available from the lab
  homepage that details the calculation of these
  rates.

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4 Figures

• x-axis Time pH
• y-axis Absorbance
• x-axis Time Temp.
• y-axis Absorbance
• x-axis pH
• y-axis Rxn rate
• x-axis Temp.
• y-axis Rxn rate

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Lab report 1 (50 points)

• Refer to Appendix A - pp. 191-195 guidelines for
  writing a core lab report.
• Ecology has all journal articles prior to 1996
  on-line. http//www.jstor.org/journals/00129658.ht
  ml

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Figures

• These take some time to get right.
• Pay close attention to format.

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All lab reports will be submitted to me AND
Turnitin.com ON TIME!!!Attached to hard copy
will be 4 hand drawn figures and signed academic
honesty form.
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• Four graphs for this lab report
• 2 figures (Absorbance vs. time)
• temperature vs. time
• pH vs. time
• 2 figures (RR vs. Tº/pH)
• Reaction rate vs temperature
• Reaction rate vs. pH
• Best fit curve for each set of data

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For this first report, ALL FIGURES MUST BE DONE
BY HAND (not by computer). In the future, you
may use computer generated figures
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• (5) English
• (3) Spelling, grammar, clarity
• (2) Format
• (1) Title
• (6) Abstract
• (12) Introduction
• (8) Background info
• (4) Hypotheses
• (6) Materials and Methods
• (4) Experimental design and materials
• (2) Data collection and analysis
• (8) Results
• (4) Tables and figures
• (2) Stats
• (2) Text
• (10) Discussion
• (8) Interpretation of data
• (2) Alternative hypothesis/sources of error
• (2) Literature cited

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For next class

• Quiz
• Bring charts to class or my office hours for me
  to look over!
• Read Ch 9 10 and Animal Taxa worksheet
• Bring dissection kits!