Biology 1 Laboratory: Sam Kosmacki

Laboratory 1 Scientific Method 2

Laboratory 2 Solutions, Acids, and Bases 5

Laboratory 3 Diffusion and Osmosis 8

Laboratory 4 Enzymes: Factors Affecting the Rate of Activity and Spectrophotometry 15

Laboratory 5 Respiration 20

Laboratory 6 The Microscope 23

Laboratory 8 Bacteria - Zone of Inhibition 28

Laboratory 9 Isolation of DNA 31

Laboratory 10 Mendelian Genetics 34

Laboratory 11 Brine Shrimp Development 1 38

Laboratory 12 Brine Shrimp Development 2 39

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Laboratory 1 Scientific Method
Purpose of Lab / Objectives / Hypothesis:
● Understand the logic and sequence of the scientific method.
● Develop productive observations, questions, and hypotheses about the natural world.
● Calculate the range, mean, and standard deviation for a set of replicate measurements.
● Design and conduct a controlled experiment to test a null hypothesis.
● Understand the difference between a hypothesis and a scientific theory.
● The further away from the sidewalk, the larger the growth of the clover.

Pre-Lab Lecture Notes:

● Science- the orderly process of posing and answering questions about the natural world
through repeated and unbiased experiments and observations
● Scientific method- an organized sequence of steps that maintains as much objectivity
and repeatability as possible
○ Make insightful observations
○ Pose and clarify testable questions
○ Formulate hypotheses
○ Do experiments to gather data
○ Quantify the data
○ Test the hypotheses
○ Refine the hypotheses and retest
○ Answer the questions and make conclusions
● Hypothesis- a statement that clearly states the relationship between biological variables,
educated guess (what you think the answer is to your question), tentative explanation for
an observation
● Null hypothesis (H0)- states that there is no difference
● Replications- repeated measures of each treatment under the same condition
● Difference between hypothesis and theory (voice thread)
● Independent variable- what you are controlling and manipulating
● Dependent variable- what you are measuring and don’t know the answer to
● Control treatment- baseline, normal
○ Controls verify that the biological response we measure is a function of the
variable being investigated and nothing else
● Null hypothesis- hypothesis of no change
● Range- difference between smallest and largest value
● Standard deviation- value that describes the variation of data points around the mean
○ smaller=higher precision
● Always find mean AND standard deviation
○ Can’t just look at the means because they may be the same, but standard
deviation could be different
● Two means are significantly different if the means plus or minus ½ of the standard
deviation do not overlap
● Do not have hypotheses that ask “why?”

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 ● Scientific questions
○ Have real answers
○ Are testable
○ Have a hypothesis that is falsifiable - your experiment could show that your
hypothesis is false (and that’s okay)
○ Are interesting
● Theory- widely accepted, extensively and rigorously tested, and exceptionally well-
supported by the data
● Never use the word prove
● Data- recorded observations or items of information
○ Qualitative- descriptions
○ Quantitative (or empirical)- recorded measurements
● Accuracy- refers to how closely the measured values agree with the true or correct value
● Precision- refers to how closely the measurements agree with each other
● Density- mass per unit of volume
● Mean- average of a group of measurements
● Median- the middle value that divides the set of measurements into two subsets of equal
size

Detailed Procedure:
Procedure 1.1 - Make insightful observations:
Observation 1: Fungi often grow on leftover food
Observation 2: Fungi such as mold and yeast grow more on leftover bread than on
leftover meat
Which of the above observations is more useful for further investigation? Why?
Observation 2 because it included more detail.

Procedure 1.2 - Posing and refining questions:

1. Question 1: Do songbird populations respond to the weather?
Question 2: Do songbirds sing more often in warm weather than in cold weather?
Which of those questions is more useful for further investigations? Why?
Question 2 because it asks the question in more detail, pertaining to more specific types
of weather. What does it mean by “respond?”
Null hypothesis- do not sing more often in certain weather
Hypothesis- sing more often in warm weather
2. General questions: What influences the distribution of pillbugs?
Specific Question 1: Does the consumption of fungi growing on leaves influence the
distribution of pillbugs?
Specific Question 2: What is the effect on the distribution of pillbugs when the pillbugs
eat leaves?

Procedure 1.3 - Formulating hypotheses:

1. Hypothesis 1: Songbirds sing more when the weather is warm.

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 Hypothesis 2: The number of bird songs heard per hour during daylight temperatures
above 80 is not significantly different from the number heard per hour at temperatures
below 80.
Hypothesis 2 is more useful- more detailed
Null hypothesis- there is no difference, no change in number of songs heard

Question 6:
a. Does the mean always describe the “typical” measurement? Why or why not?
No because the mean takes into account outliers on both ends of the spectrum.
b. What information about a sample does a mean not provide?
Mean does not provide a range of numbers.
Question 7:
a. What is responsible for this difference between the mean and median
The really low number would cause the mean to drop more than the median-
outliers: variable that is far off from the other precise numbers.
b. How would the median change if the 9-mm-long leaf was not in the sample?
It would not change. The median remains 64.
c. How would the mean change if the 9-mm-long leaf was not in the sample?
The mean would increase because the value of 9 was an outlier on the lower end
of the data set.
d. What is the mean for sample 1? 30
What is the mean for sample 2? 30
Question 8:
a. Could two samples have the same mean but different ranges? Explain.
Yes, they could have the same mean but different ranges. The range is the
highest value minus the lowest value. The mean is the average of all of the values. The
range may vary between the two sets because the highest and lowest values are
different. Despite this, the values in the two data sets could still give you the same
means. This would be dependent on the number of values and what those values are.
b. Could two samples have the same range but different means? Explain.
Yes, they could have the same range but different means. The range is the
highest value minus the lowest value. The mean is the average of all of the values. The
range of two sets could equal the same value, but if the values in between vary from
data set to data set, the mean would differ. Additionally, there could be a different
number of values for each data set, which would also affect the mean.

Observations:
● Closer to sidewalk: 1.9 cm, 1.5 cm, 2.5 cm
○ Group of 3 clovers about 1 foot from the sidewalk
● Further from sidewalk: 3.0 cm, 4.1 cm, 5.5 cm, 7.1 cm, 8.0 cm
○ Group of 9 clovers

Data:

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 ● 1.5 cm, 2.5 cm, 3.0 cm, 4.1 cm, 5.5 cm, 7.1 cm, 8.0 cm
● Mean = 4.2 cm
● Median = 3.6 cm
● Range = 6.5 cm
● Standard deviation = 2.4
● Variance = 5.9

Conclusions / Summary:
The closer to the sidewalk the clovers grew, the shorter and less abundant they were. The
further from the sidewalk the clovers grew, they were able to grow taller and in more
abundance.

Laboratory 2 Solutions, Acids, and Bases

Purpose of Lab / Objectives / Hypothesis:
● Calculate correct ratios of, and prepare, percent solutions.
● Calculate and prepare dilutions.
● Demonstrate proper pipetting technique.
● Measure the pH of various liquids.
● Demonstrate that buffers stabilize the pH of a liquid.

Pre-Lab Lecture Notes:

● Solution = solute + solvent
● Concentrations often expressed as percentage or molarity
● 1 mole = molecular weight (grams)
○ Molecular weight g/mol
● Molarity (M) = moles solute / liters of solution
● Stock solution = concentrated form
○ Want various concentration levels that you can dilute and make very easily from
your stock
● Dilution equation:
○ C (original) x V (original) = C (final) x V (final)
○ C = concentration, V = volume
○ Don’t need molecular weight for dilution
○ Take final mL - original to determine how much water to add
● Mole- standard measure of the amount of a chemical
● Acids- molecules that release hydrogen ions when dissolved in water
● Bases- molecules that remove hydrogen ions from solution

Detailed Procedure:
Procedure 5.1 - Measure the pH of liquids:
300 microliters

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Apple cider Vinegar- 4
1% milk- 6
Sprite- 3
Soap solution- 6

Question 1:
a. How many grams of sucrose would you dissolve in water for a total volume of 500 mL to
make a 5% (weight/volume) solution?
mass(g)
% concentration= x 100
volume (mL)
mass( g)
5 %= x 100
500mL
mass=25 g
b. How many grams of calcium chloride would you add to water for a total volume of 500
mL to make a 5% (weight/volume) solution?
mass(g)
% concentration= x 100
volume (mL)
mass( g)
5 %= x 100
500mL
mass=25 g
c. How many grams of calcium chloride would you add to water for a total volume of 100
mL to make a 5% (weight/volume) solution?
mass(g)
% concentration= x 100
volume (mL)
mass( g)
5 %= x 100
100mL
mass=5 g

Question 2:
a. How many grams of NaCl (molecular weight = 58.5 g/mol) would you dissolve in water to
make a 0.5 M NaCl solution with 500 mL final volume?
mol g
0.5 x 58.5 x 0.5 L=14.6 g
L mol
b. How many grams of NaCl (molecular weight = 58.5 g/mol) would you dissolve in water to
make a 50 mM NaCl solution with 500 mL final volume?
mol g
0.05 x 58.5 x 0.5 L=1.46 g
L mol
c. How many grams of sucrose (molecular weight = 342 g/mol) would you dissolve in water
to make a 0.22 M sucrose solution with 1 L final volume?
mol g
0.22 x 342 x 1 L=75.2 g
L mol

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 d. How many grams of sucrose (molecular weight = 342 g/mol) would you dissolve in water
to make a 0.22 mM sucrose solution with 100 mL final volume?
mol g
2.2 x 1 0−4 x 342 x 0.1 L=7.52 x 1 0−3 g
L mol
e. How many grams of calcium chloride (molecular weight = 111 g/mol) would you dissolve
in water to make a 0.111 M solution with 1 L final volume?
mol g
0.111 x 111 x 1 L=12.3 g
L mol
f. How many grams of calcium chloride (molecular weight = 111 g/mol) would you dissolve
in water to make a 0.2 M solution with 200 mL final volume?
mol g
0.2 x 111 x 0.2 L=4.4 g
L mol
g. If you were presented with 2 L of a 2 M sucrose stock solution, how many grams of
sugar would be in a 100 mL aliquot?
mol g
2 x 342 x 2 L=1368 g
L mol
2 L÷ 0.1 L=20
1368 g÷ 20=68.4 g
68.4 g

Question 3:
a. How many milliliters of concentrated (18 M) sulfuric acid are required to prepare 750 mL
of 3 M sulfuric acid?
V i Mi = V f M f
V i (18 M )=(750 mL)(3 M )
V i=125 mL
b. How would you prepare 100 mL of 0.4 M magnesium sulfate from a stock solution of 2 M
magnesium sulfate?
V i Mi = V f M f
V i (2 M )=(100 mL)(0.4 M )
V i=20 mL
You need 20 mL of the stock solution to prepare 100 mL of magnesium sulfate.

c. How many milliliters of water would you add to 100 mL of 1.0 M HCl to prepare a final
solution of 0.25 M HCl?
V i Mi = V f M f
(100 mL)(1.0 M )=V f ( 0.25 M )
V f =400 mL
400 mL−100 mL=300 mL
300 mL

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Observations:

Data:
● 300 microliters
● Apple cider Vinegar- pH of 4
● 1% milk- pH of 6
● Sprite- pH of 3
● Soap solution- pH of 6

Conclusions / Summary:
The pH of apple cider vinegar, 1% milk, sprite, and soap solution were all acidic. Sprite
was the most acidic with a pH of 3 and milk and the soap solution were the least acidic with a
pH of 6. Apple cider vinegar was a little less acidic than the sprite was, with a pH of 4.

Excellent job on your lab notebook so far! Glad to see that you have already started this week’s
entry!

Laboratory 3 Diffusion and Osmosis

Purpose of Lab / Objectives / Hypothesis:
● Explain the factors controlling a substance’s direction and rate of diffusion.
● Predict the direction and rate of osmosis into and out of cells surrounded by hypotonic,
hypertonic, and isotonic environments.
● Describe how hypotonic, hypertonic, and isotonic solutions affect the volume and
integrity of blood cells.

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 ● Perform and interpret qualitative tests to detect the presence of biologically important
macromolecules.
● Explain the importance of a positive and a negative control in a biochemical test.
● Prediction:
○ Bag A: from outside to inside of the bag
○ Bag B: isotonic - no movement
○ Bag C: from inside of the bag to the outside
○ Bag D: from inside of the bag to the outside

Pre-Lab Lecture Notes:

● 4 main macromolecules / organic compounds are carbohydrates, proteins, lipids, and
nucleic acids (biomolecules)
● Polymers made of linked monomers
● Dehydration synthesis- an energy-requiring process in which a molecule of water is
removed and the 2 subunits are bonded covalently, links subunits of macromolecules
● Hydrolysis- energy-releasing process in which breaking the bond between the subunits
requires the addition of a water molecule and releases energy
● Biochemical tests require at least 2 treatments, an unknown solution to be identified and
controls to provide standards for comparison
● Positive control- variable for which you are testing; reacts positive and demonstrates the
test’s ability to detect what you expect
● Negative control- does not contain the variable for which you are searching, only the
solvent (usually distilled water) and does not react in the test
● Qualitative tests- absence or presence of particular substance in the sample
● Monosaccharides- simple sugars (ex: carbohydrates)
● Disaccharide- paired monosaccharides (ex: sugar = glucose+fructose)
● Polysaccharides- linking at least 3 monosaccharides (ex: starch, glycogen, cellulose)
● Reducing sugars- process free aldehyde or ketone groups that reduce weak oxidizing
agents such as the copper in Benedict’s reagent
● Benedict’s reagent- contains cupric (copper) ion complexed with citrate in alkaline
solution
○ Identifies reducing sugars based on their ability to reduce the cupric ions to
cuprous oxide (Cu2+ to Cu+)
● Green solution indicates a small amount of reducing sugars, reddish orange indicates an
abundance of reducing sugars
● Proteins are versatile structural molecules found in all life forms
● Peptide bond- forms between the amino group of one amino acid and the carboxyl group
of an adjacent amino acid and is identified by a Biuret test
● Lipids include molecules that dissolve in nonpolar solvents (acetone, methanol, or
ethanol), but not well in polar solvents (water)

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 ○ Tests are based on the ability to absorb pigments in fat-soluble dyes
● Triglycerides are abundant lipids made of glycerol and 3 fatty acids
● DNA and RNA are nucleic acids made of nucleotide subunits
● Dische diphenylamine test- identifies DNA in acidic conditions (intensity of color is
proportional to concentration)
● Heat causes random motion of molecules that passively moves molecules in biological
systems
● Brownian movement- can see small particles move after they collide with moving
molecules
● Diffusion- passive (no energy required), directional movement of molecules, move from
areas of high concentration, heat, and pressure to an are of low concentration, heat, and
pressure
○ Determined by steepness of gradient, molecular size, polarity, or solubility
● Differentially permeable- allow passage of small molecules, not bigger ones
● Membrane permeability depends on solute size, charge, polarity, lipid solubility (polar vs
nonpolar)
● Dialysis- separation of dissolved substances by means of their unequal diffusion through
a differentially permeable membrane (model)
● Phenolphthalein- pH indicator that turns red in basic solutions
● Iodine- starch indicator that changes from yellow to dark blue in presence of starch
○ Distinguishes starch from other saccharides
● Osmosis- diffusion of water across a semipermeable membrane
● Solution- homogeneous, liquid mixture of 2 or more kinds of molecules
● Solvent- dissolves substances, solute- substance dissolved in a solution
● Hypotonic- solution with a lower concentration of solutes, water will flow into cell,
swells, could potentially burst
● Hypertonic- solution with a higher concentration of solutes (outside of the cell), water
will want to flow out of the cell to equal the concentration gradient, cell may shrivel
● Isotonic- equal concentrations inside and outside of cell

Detailed Procedure:
Procedure 9.4 - Observe osmosis across a concentration gradient:

Table 9.1
Changes in Weight of Dialysis Bags Used as Cellular Models*

0 Min 10 Min 20 Min 25 Min

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 Initial Total Change Total Change Total Change
Weight Weight in Weight in Weight in
Weight Weight Weight

Bag A 9.52 g 9.33 g 0.19 g 9.17 g 0.16 g 8.99 g 0.18 g

Bag B 6.36 g 6.44 g 0.08 g 6.48 g 0.04 g 6.51 g 0.03 g

Bag C 8.00 g 8.23 g 0.23 g 8.65 g 0.42 g 8.63 g 0.02 g

Bag D 5.61 g 6.24 g 0.63 g 6.61 g 0.37 g 6.76 g 0.15 g

5.)
a. Did water move across the membrane in all bags containing solutions of sugar?
No, water did not move across the membrane in all bags containing solutions of
sugar.

b. In which bags did osmosis occur?

Osmosis occurred in bags A, C, and D.

c. A concentration gradient for water must be present in cells for osmosis to occur. Which
bag represented the steepest concentration gradient relative to its surrounding
environment?
Bag D represented the steepest concentration gradient. The first change in
weight after 10 minutes was 0.63 grams. Then the next change in weight after another 10
minutes was 0.37 grams. The final change in weight after an additional 5 minutes was 0.15
grams.

d. The steepest gradient should result in the highest rate of diffusion. Examine the data in
table 9.1 for Change in Weight during the 15- and 30-minutes intervals. Did the greatest
changes in weight occur in cells with the steepest concentration gradients? Why or why
not?
Yes, the greatest changes in weight occur in cells with the steepest concentration
gradients. This is because the greater the difference in weight, the steeper the concentration
gradient will be. These are more drastic changes. This is also a result of the higher
concentrations of sucrose.

Procedure 6.1 - Perform the Benedict’s test for reducing sugars:

Table 6.1

Solutions and Color Reactions for Benedict’s Test for Reducing Sugars

Tube Solution Benedict’s Solution Benedict’s Color

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 Reaction

1 10 drops potato juice 3 mL Hot: blue

Room Temperature:
blue

2 10 drops onion juice 3 mL Hot: green (cloudy)

Room Temperature:
darker cloudy green

3 10 drops sucrose 3 mL Hot: blue

solution Room Temperature:
blue

4 10 drops glucose 3 mL Hot: blue

solution Room Temperature:
blue

5 10 drops reducing- 3 mL Hot: orange (cloudy)

sugar solution Room Temperature:
darker cloudy orange

6 10 drops starch 3 mL Hot: blue

solution Room Temperature:
blue

7 10 drops distilled 3 mL Hot: blue

water Room Temperature:
blue

2.
a. Which of the solutions is a positive control? Negative control?
The positive controls were the onion juice, which turned green, and the
reducing-sugar solution, which turned orange. The negative controls were the potato juice,
sucrose solution, glucose solution, starch solution, and distilled water, which all remained blue.

b. Which is a reducing sugar, sucrose or glucose? How do you know?

Glucose is a reducing sugar. Although it did not completely change color,
the solution was a slightly darker shade of blue than the sucrose solution. The glucose solution
should have tested positively, while the sucrose solution tested negatively, with no change in
color solution.

c. Which contains more reducing sugars, potato juice or onion juice? How do you
know?
The onion juice contains more reducing sugars. I know this because the
solution with the onion juice turned green, while the solution with the potato juice remained blue.

d. What does this tell you about how sugars are stored in onions and potatoes?

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 Since the onion juice tested positively, this tells you that sugars are more
concentrated in onions. This would also tell you that there is more energy stored in onions than
in potatoes.

Procedure 9.5 - Graph osmosis:

Observations:

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Data:
● Bag A (10 mL of 1% sucrose solution in 10% sucrose solution):
○ Initial weight- 9.52 g
○ Weight after 10 minutes- 9.33 g
○ Weight after another 10 minutes- 9.17 g
○ Weight after another 5 minutes- 8.99 g
● Bag B (10 mL of 1% sucrose solution in 1% sucrose solution):
○ Initial weight- 6.36 g
○ Weight after 10 minutes- 6.44 g
○ Weight after another 10 minutes- 6.48 g
○ Weight after another 5 minutes- 6.51 g
● Bag C (10 mL of 10% sucrose solution in 1% sucrose solution):
○ Initial weight- 8.00 g
○ Weight after 10 minutes- 8.23 g
○ Weight after another 10 minutes- 8.65 g
○ Weight after another 5 minutes- 8.63 g
● Bag D (10 mL of 20% sucrose solution in 1% sucrose solution):
○ Initial weight- 5.61 g
○ Weight after 10 minutes- 6.24 g
○ Weight after another 10 minutes- 6.61 g
○ Weight after another 5 minutes- 6.76 g

Conclusions / Summary:
For procedure 9.4, osmosis occurred in bags A, C, and D. Bag D had the steepest
concentration gradient because it had the biggest changes in weight throughout the experiment.
The greater the difference in weight, the steeper the concentration gradient will be. For
procedure 6.1, the onion juice solution turned green and the reducing-sugar solution turned
orange, demonstrating that they were the positive controls. All of the other solutions remained
blue, thus making them the negative controls. Since the onion juice was a positive control, while
the potato juice was a negative control, we know that onion juice contains more reducing sugars
and sugars are more concentrated in onions than in potatoes.

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Laboratory 4 Enzymes: Factors Affecting the Rate of
Activity and Spectrophotometry
Purpose of Lab / Objectives / Hypothesis:
● Describe the relationship between structure and function of enzymes.
● Relate structure and function to active sites and optimal conditions for enzymatic activity.
● Hypothesize and test how changes in temperature and pH affect enzymatic reaction
rates.
● Describe how some enzymatic reaction rates can be measured by color changes and
gas liberation as products are formed.
● Operate a spectrophotometer.

Pre-Lab Lecture Notes:

● Enzymes- speed up chemical reactions, allow chemical reaction to take place more
readily, reduces activation energy needed
● Active site bonds to substrate (molecule enzyme reacts on), form enzyme-substrate
complex
● Enzymes in system are continuously reused
● Enzymes can denature if in too high of temperature
● Spectrophotometry- put solution into sample holder, shines beam of light at sample and
measures the amount of light that comes through/ is transmitted
○ Measures light as a specific wavelength
○ Each has a fingerprint of wavelengths it will absorb or reflect
■ Can determine molecules based on this
○ Can measure concentrations
○ Blank is a test tube absorbs only chemical you are absorbing(sample of just the
solvent), contains everything except for the analyte you are interested in
observing; gives accurate reading
■ Put it in spectrophotometer before analyzing solutions
● pH affects enzyme activity
○ As pH lowers becomes more acidic, side chain gains hydrogen ions
○ Very extreme pH could change enzyme shape and will render ineffective
● Inhibitors affect enzyme activity
● Competitive inhibition- competes with enzyme for substrate, slows down chemical
reaction, using up all the substrate and enzymes can’t get to it
● Enzymes are biocatalysts
● Biocatalysts- accelerate metabolic reactions to biologically useful rates

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 ● Enzyme-substrate complex stresses/distorts chemical bonds and forms a transition state
in which the substrate becomes more reactive
● Energy of activation- energy needed to form the transition state, is lowered by the
enzyme
● Active site- site of attachment and the surrounding parts of the enzyme that stress the
substrate’s bonds
● Reaction is complete when the product forms and the enzyme is released in its original
condition
● Enzymes are reusable
● See formula on page 114
● Denature- destroy effectiveness of enzyme by altering active site and slowing the
reaction
● Optimal conditions- the range of values for environmental factors such as temperature
and pH within which an enzyme functions best
● Catechol oxidase- a plant enzyme that converts catechol to benzoquinone, effects of
temperature on enzyme activity can be investigated with this (see equation page 115)
● When fruit is bruised, injured cells release catechol and catechol oxidase, which react to
from brownish product
● See diagram page 81
● Spectrophotometry- one of our most versatile and precise techniques for assays ranging
from blood chemistry to pollutants in water
● Light source of a spectrophotometer produces white light, a combination of all visible
wavelengths
● Filter is adjusted to select the wavelength that you wish to pass through the sample
● The sample is a solution contained in a clear test tube or cuvette made of glass or quartz
● A blank is a test tube or cuvette containing only the solvent used to dissolve the
chemical you are analyzing
● Photodetector converts light energy into electrical energy
● The meter on the front of the spectrophotometer measures the electrical current
produced by photodetector and displays the results on a scale of absorbance or
transmittance values
● Absorbance- amount of radiation retained by the sample
● Transmittance- amount of radiation passing through the sample
● Test tubes 1-3 act as controls
● Test tube 3 is going to act as our blank (setting up baseline, measures 0 point, then
when enzyme is added, what difference does it make)
● Competitive inhibition- molecules structurally similar to the substrate and therefore
competitive for positions at the active sites of enzymes

Detailed Procedure:
Procedure 11.1 Determine the effect of temperature on catechol oxidase activity-

Table 11.1

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 Tube Distilled pH 6 Buffer Potato 1% Catechol Temperature
Water Extract
(catechol
oxidase)

1 2 mL 1 mL 22ºC

2 1 mL 1 mL 1 mL 22ºC

3 1 mL 1 mL 1 mL 22ºC

4 1 mL 1 mL 1 mL 22ºC

5 1 mL 1 mL 1 mL 4ºC

6 1 mL 1 mL 1 mL 40ºC

7 1 mL 1 mL 1 mL 80ºC

Hypothesis: The second lowest temperature will cause the most optimal results.

Table 11.2

Qualitative and Quantitative Color Changes as Catechol Oxidase Activity Produces

Brown Benzoquinone

Qualitative Color Change Results Quantitative Absorbance Results

Tube 0 min 5 min 10 min 15 min 0 min 5 min 10 min 15 min

1 0 0 0 0 0 0 0 0

2 0 0 0 0 70 58 45 50

3 0 0 0 0 0 0 0 0

4 2 1 1 1 70 30 33 34

5 2 2 2 2 59 27 29 26

6 1 1 2 2 54 31 25 26

7 1 1 2 2 54 42 33 43

Qualitative Color Change Key:

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No change 0
Little more change 1
Moderate change 2
Dark color change 3

1.
a. Do your data support or refute your hypotheses?
My data supports my hypothesis because the tube placed in the 22oC
temperature (the second lowest temperature) demonstrated the most
change, and thus the most optimal results.
b. Write a hypothesis and null hypothesis for the effect of temperature on catechol
oxidase activity.
Hypothesis: Room temperature environments provide the greatest the
catechol oxidase activity.
Null hypothesis: The higher the temperature of the environment, the
greater the catechol oxidase activity.
c. What were the enzyme, substrate, and product of the enzymatic reaction?
The enzyme was the catechol oxidase (potato extract), the substrate was
the 1% catechol, and the product of the reaction was benzoquinone.
d. Why was each tube left undisturbed for 5 min in step 6 of procedure 11.1?
The tubes were left undisturbed for 5 minutes in order to let the liquid
change to room temperature.
e. Explain the results observed for tubes 1-3. What was the purpose of these
tubes?
Tube 2’s quantitative results were 70, 58, 45, and 50 and there was no
visible change in the liquid. Tubes 1 and 3 demonstrated no change. The
purpose of these tubes was to act as controls for the experiment. Tube 1
was the control for the distilled water, tube 2 was the control for the 1%
catechol, and tube 3 was the control for the substrate. Additionally, tube 3
acted as the blank, which helped us set up the baseline and was the
sample with just the solvent.

f. Graph of tubes 4-7

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Figure 1. The enzyme activity of catechol oxidase under different temperatures for four tubes.

Observations:
● No color change for control tubes (tubes 1-3)
● Tubes 4-7 all demonstrated color change
● Tube 4 demonstrated the most change quantitatively
● Tube 5 demonstrated the most change qualitatively

Data:
● Tube 2: 70, 58, 45, 50; 0, 0, 0, 0
● Tube 4: 70, 30, 33, 34; 2, 1, 1, 1
● Tube 5: 59, 27, 29, 26; 2, 2, 2, 2
● Tube 6: 54, 31, 25, 26; 1, 1, 2, 2
● Tube 7: 54, 42, 33, 43; 1, 1, 2, 2
Conclusions / Summary:
Tube 4, which had the pH 6 buffer, potato extract, and 1% alcohol in 22oC, presented
with the most change, both qualitatively and quantitatively. This was the tube placed in the room
temperature and showed the most change. All of the tubes that included 1% catechol, which
was all of them besides tubes 1 and 3, presented some sort of change.

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Laboratory 5 Respiration
Purpose of Lab / Objectives / Hypothesis:
● List and describe the processes of aerobic and anaerobic respiration.
● Explain the function and importance of respiration to life.
● Describe fermentation and the two different byproducts produced.
● List the economic importance of fermentation.
● Measure carbon dioxide as a product of fermentation.

Pre-Lab Lecture Notes:

● Cellular respiration- oxidation of organic molecules and a concomitant release of energy
● Some of that energy stored in chemical bonds of ATP, which is used as source of
energy for cellular metabolism
● Glycolysis- oxidation of glucose to pyruvate, some of the energy released from each
glucose molecules is stored in ATP, occurs with or without oxygen
● Krebs cycle- when oxygen is present, organisms continue respiration by oxidizing
pyruvate to CO2
● Aerobes- organisms that use oxygen for respiration beyond glycolysis
● Electron transport chain- generates proton gradients from energy stored in reduced NAD
and related compounds that lead to formation of approximately 18-times more ATP that
formed in glycolysis
● Oxygen is the final electron-acceptor and is reduced from H2O
● See equation on page 126
● Anaerobes- live without oxygen and may even be killed by oxygen in the atmosphere
● Anaerobic fermentation- reduces pyruvate from glycolysis to either CO2 and ethanol or
lactic acid
○ ATP and heat are also products of anaerobic fermentation
● See equations on page 127
● Anaerobic fermentation produces 18-fold less ATP per glucose molecule than does
aerobic respiration
● Pyruvate- a product of glycolysis
● Magnesium sulfate- provides Mg2+, a cofactor that activates some enzymes of glycolysis
● Sodium fluoride- an inhibitor of some enzymes of glycolysis
● Glucose- a common organic molecule used as an energy source for respiration
● Carbon dioxide and water produce carbonic acid
● You can monitor cellular respiration by measuring acid production as change in pH
● pH- measure of the acidic or basic properties of a solution
● The rate of O2 uptake during cellular respiration indicates the metabolic rate of an
organism
● Aerobic respiration: glucose and oxygen yield carbon dioxide, water, ATP, and heat
● Economic applications
○ Fermentation- bread, alcohol, pickled foods (lowers pH, organisms that can make
food go bad are not present in that environment)

20
 ● Test the effects of temperature on fermentation
● Measure CO2 production, by measuring circumference of the balloons

Detailed Procedure:
● Retrieve four test tubes and place them into a test tube rack
● Fill test tubes about ¾ full with distilled water
● Measure 1 gram of yeast in a weigh boat, repeat this four times and transfer into each of
the test tubes
● Repeat previous step with glucose
● Use stir rod to mix samples
● Place balloon over the top of all four tubes
● Place each tube under different temperature conditions; 60°C, 30°C, room temperature
(22°C) and fridge temperature (4°C)
● Measure the circumferences of the balloons by wrapping string around the balloon and
measuring the length of the string (in centimeters) with a ruler
● Recorde date in data table and create a graph

1. Why must organisms such as yourself inhale oxygen and exhale CO2?
a. Organisms must inhale oxygen because it is the final electron acceptor in the
electron transport chain. Without oxygen this chain will not be functional and the
anaerobic organism will die. We must exhale CO2 because it is a waste product
of respiration that must be released.
2. What are the advantages and disadvantages of anaerobic fermentation?
a. Some advantages are that fermentation can occur even if there is not a sufficient
amount of oxygen, it is a fast process, and is a relatively simple process. The
major disadvantage is that fermentation does not produce a lot of ATP, 18-fold
less per glucose molecule than aerobic respiration.

Observations:
● Balloons in cooler temperatures showed little to no change in inflation / circumference
● Balloons in warmer temperatures showed inflation

Data:

Rate of Fermentation Under Different Temperature Conditions

Temperature (°C) Circumference (cm)

21
 4 10.5 (no inflation)

22 10.6

37 11.8

60 15.0
Figure 1. Chart showing the rate of fermentation within the four concentrations under different

temperature conditions.

Figure 2. Line graph showing the change in balloon circumference according to rise in

temperature.

Conclusions / Summary:
Based on our experimental results, our hypothesis was supported. As the temperature of

the sample increased, the rate of enzyme activity increased, and as a result, the rate of

fermentation also increased. The tubes put under colder temperatures had little to no change in

circumference, leaving the balloon looking deflated. The tube left in the fridge at 4°C showed no

22
change in circumference staying at 10.5 centimeters. The tube left at room temperature (22°C)

showed a small change with a circumference of 10.6 centimeters. The tubes left in warmer

temperatures however, showed greater changes in circumference which resulted in the balloons

looking inflated. The tube left in the first hot water bath at 37°C had a circumference of 11.8

centimeters. The second tube left in the second hot water bath at 60°C had a circumference of

15.0 centimeters which was the greatest increase. These results align with the ideas in our

hypothesis that a higher temperature will increase the enzyme activity and rate of fermentation.

This causes carbon dioxide to be released and fill up the balloons. Additionally, although we did

not test a temperature high enough to see the denaturing of enzymes, we can predict that once

the temperature gets too hot, enzymes will denature, the rate of fermentation will decrease, and

the balloons would not fill up with as much carbon dioxide.

Laboratory 6 The Microscope

Purpose of Lab / Objectives / Hypothesis:
● Identify and explain the functions of the primary parts of a compound and a dissecting
microscope.
● Properly transport, focus, and use a compound and dissecting microscope to examine
biological specimens.
● Prepare a wet mount, determine magnification and size of the field of view, and
determine the depth of view.
● Use the compound microscope with a variety of prepared slides.

Pre-Lab Lecture Notes:

● Light microscope- coordinated system of lenses arranged to produce an enlarged,
focusable image of a specimen, magnifies a specimen
● Resolution- ability to distinguish two points as separate points, makes image look
sharper
● Contrast- the difference between the lightest and darkest parts of an image
○ Dyes are often used to increase contrast and make the specimen more visible
● The light microscope has three systems: illuminating, imaging, and sometimes a viewing
and recording system

23
 ● Illuminating: concentrates light on the specimen, consists of light source, condenser
lens, and iris diaphragm
○ Light source- lightbulb located at the base of the microscope, passes light
through thin part of specimen
○ Condenser lens- located immediately below the specimen, focuses light from the
light source onto the specimen
○ Condenser iris diaphragm- below condenser, knurled ring or lever that can be
opened and closed to regulate the amount of light reaching the specimen, image
is bright when open and dim when closed
● Imaging: Ocular- lense that you look through
○ Monocular and binocular microscopes
○ Magnify image 10 times
● Body tube- metal casing through which light passes to the ocular, contains mirrors and a
prism that redirect light to the oculars
● Stage- secures the glass slide on which the specimen is mounted
● See image page 23
● Viewing and recording system: if present, converts radiation to a viewable and/or
permanent image, consists of camera or video screen
● Dissecting (stereoscopic) microscope- offers some advantages over a compound
microscope, much larger working distance, use light source above specimen (helpful for
thicker specimens), image is from reflecting light
○ Always binocular, 3-D image
○ Disadvantage- lower resolution and magnification than compound microscope
● Working distance- distance between the objective lens and specimen
● Start with low stage (stage where you place slide should be pushed down as far as
possible) and lowest objective power (4x)
● Field of view- area you can see without moving the slide or stage, gets smaller with
increasing power
● Measure in 4x in mm
● Only need to measure FOV for each objective 1x

Detailed Procedure:
● This lab was all online
● Procedure 3.1

1.
a. As you view the letter e, how is it oriented? Upside down or right side up?
Did not complete the lab in class.
b. How does the image move when the slide is moved to the right or left? Toward
you or away from you?
Did not complete the lab in class.
c. What happens to the brightness of the view when you go from 4x to 10x?
Did not complete the lab in class.
● Procedure 3.2

24
Total Magnifications and Areas of Field of View (FOV) for Three Objectives

Objective Objective Ocular Total FOV FOV Measurement

Power Magnification Magnification Magnification Diameter Area (mm) for 1
x = (mm) (mm2) Ocular Space

4x x =

10x x =

40x x =

2.
a. How many times is the image of the e magnified when viewed through the high-
power objective?
40 x 10 = 400x
b. If you didn’t already know what you were looking at, could you determine at this
magnification that you were looking at a letter e? How?
Yes, you could move the slide around to view the different parts of the
image until you are able to determine that it is the letter e.
● Procedure 3.3- Alternate procedure: use a transport ruler to determine the size of the
field of view (page 27)
3.
a. Which provides the largest field of view, the 10x or 40x objective?
10x because it is less magnified than 40x.
b. How much more area can you see with the 4x objective than with the 40x
objective?
3.0mm x 4 = FOVhigh x 40
FOVhigh = 0.30 mm
3.0 mm - 0.30 mm = 2.7 mm
c. Why is it more difficult to locate an object starting with the high-power objective
than with the low-power objective?
It is more difficult because you are starting at a greater magnification.
This means that you may not be able to see the full specimen you are
viewing. If you start at the low-power objective you can focus on a larger
field of view and then increase the magnification and refocus again,
allowing you to better view the specimen.
d. Which objective should you use to initially locate the specimen? Why?
You should use the low-power objective to initially locate the specimen
because it shows a larger field of view and allows you to see more of the
specimen. You can then increase objectives and refocus to see the specimen
more closely.
● Procedure 3.4 depth of field

25
 ○ Thickness of object only one thread is focus at a time, large- all threads are in
focus (how does the depth of field change with magnification?)
4.
a. Are all three colored threads in focus at low power?
Did not complete this lab in class.
b. Can all three threads be in focus at the same time using the high-power objective?
Did not complete this lab in class.
c. Which objective, high or low power, provides the greatest depth of view?
High power will provide the greatest depth of view.
● Procedure 3.5
○ Use pond water
○ 45 degree angle to minimize amount of air bubbles
5.
a. Is the image always best with the highest illumination?
No, higher illumination is better for higher power.
b. Is the same level of illumination best for all magnifications?
No, lower illumination is better for lower power and higher illumination is better
for higher power.
c. Which magnifications require the most illumination for the best clarity and contrast?
Higher objectives require more illumination.
6.
a. Why is it important to put a coverslip over the drop of water when you prepare a wet
mount? That is, what are the functions of a coverslip?
The coverslip is important so that the microscope and specimen do not touch and
so that the specimen stays in the same area on the slide.
● Procedure 3.6
7.
a. What is the area of the field of view when you use the lowest magnification of your
dissecting microscope? What about when you use the highest magnification?
Did not complete the lab in person.
b. Place a microscope slide of the letter e on the stage. As you view the letter e, how is it
oriented?
Did not complete the lab in person.
c. How does the image through a dissecting microscope move when the specimen is
moved to the right or left? Toward you or away from you?
Did not complete the lab in person.
d. How does the direction of illumination differ in dissecting as opposed to compound
microscopes?
In compound microscopes, light is transmitted from a light source below the
specimen. In dissecting microscopes, a light source above the specimen is used which
produces an image formed from reflected light.
e. What are some examples of biological specimens that would be best examined with a
dissecting microscope instead of a compound microscope?

26
 Thinner specimens that are able to allow light through them would be observed
under a compound microscope. Larger and more opaque specimens such as butterfly wings
and organisms found in pond water can be observed under a dissecting microscope.

A Comparison of Dissecting and Compound Microscope

Characteristic Dissecting Microscope Compound Microscope

Magnification

Resolution

Size of field of view

Depth of field

Observations:

Data:

Conclusions / Summary:

Laboratory 8 Bacteria - Zone of Inhibition

Purpose of Lab / Objectives / Hypothesis:
● Explain the zone of inhibition and its usefulness to everyday life.
● Set up an experiment to test various antimicrobial agents.
● Perform (virtually) and interpret a Gram stain.
● Use Microsoft Excel for graphing.
● Find primary literature pertaining to a topic and complete an annotated bibliography.

Pre-Lab Lecture Notes:

● Prokaryotes- species with cells lacking membrane-bound organelles
● Eukaryotes- those with membrane-bound organelles and include plants, animals, fungi,
and protists
● Domain Archaea includes kingdom Archaebacteria, which are all prokaryotes
● Domain Bacteria includes kingdom Bacteria, which are all prokaryotes and the most
abundant organisms on Earth
● Domain Eukarya includes kingdoms Fungi, Plantae, Animalia and the polyphyletic
(multiple origins) protists

27
● Many Archaebacteria are called extremophiles because they survive in habitats that are
often extremely acidic, hot, or salty
● Most bacteria are heterotrophic- they derive their energy from organic molecules made
by other organisms
● Heterotrophic bacteria are decomposers- they feed on dead organic matter and release
nutrients locked in dead tissue
● See images and diagrams of pages 254-256
● Autotrophic- bacteria that derive their energy from photosynthesis or the oxidation of
inorganic molecules
● Bacteria reproduce asexualy via binary fission- a cell’s DNA replicates and the cell
pinches in half without the nuclear and chromosomal events associated with mitosis
● Some bacteria have genetic recombination via conjugation- all or part of the genetic
material of one bacterium is transferred to another bacterium and a new set of genes is
assembled
● Gram stain- one of the most important techniques to classify bacteria, based on the
different structural and chemical compositions of bacterial cell walls
● Gram- positive bacteria- have a thick cell wall that retains a purple dye
● Gram-negative bacteria- have a much thinner cell wall that does not retain the dye
● Nitrogen fixation- certain bacteria and cyanobacteria transform atmospheric nitrogen into
other nitrogenous compounds that can be used as nutrients by plants
● Sensitivity plate- petri dish of solid medium that has been uniformly inoculated on its
entire surface with a known bacterium or an unknown sample from an infected patient
● After 24 hours, an effective antibiotic will produce a visible halo of clear surface around
the disks where it inhibited growth of the bacteria
● If the antibiotic was ineffective, the bacteria will grow to the edge of the paper disk
● See image on page 264
● All living things contain cells
● Eukaryotes: more than one cell
● Prokaryotes: one cell organisms
● Bacteria are prokaryotes, meaning they are only ONE celled organisms
● Bacteria are classified by:
○ Shape- spirilla (spiral), Cocci (sphere), and Bacilli (rod)
○ Gram stain- chemistry of bacterial cell wall differs between species
■ Gram-positive- thick layer, peptidoglycan, stain purple
■ Gram-negative- thinner cell wall, outer envelope of glyco polysaccharides,
turns pink, prevents from being stained
● We can grow bacteria on different types of media (medium)
● Media is a liquid or gel designed to support growth of microorganisms or cells
● Need to make sure bacteria has proper nutrients for growth
● Wear goggles and gloves, Q-tip trash in small bag at bacteria station, all other trash into
biohazard
● Goal 1: test if antiseptics or disinfectants affect different bacteria (do they kill bacteria
like they are supposed to or can bacteria still grow on them)
● Antiseptic- chemical substances that inhibit bacterial growth (applied to living tissue)

28
 ● Disinfectant- chemical substances that inhibit bacterial growth (applied to non-living
objects)
● Zone of inhibition- area where bacterial growth was inhibited
○ The bigger the zone of inhibition, the better the antimicrobial agent
● 4 test disks and 1 control
● Goal 2: find relevant primary literature, make annotated bibliography- list of citations,
followed by about 150 words (description), relevance, accuracy, and quality of source
you selected

Detailed Procedure:
● In class procedure:
○ Swab bottom of shoe with Q-tip
○ Swab the same Q-tip on the media in the dish
○ Let sit for a week and observe the growth of bacteria
● Annotated bibliography procedure:
○ Utilize EbscoHost database on Seton Hill Library
○ Peer review, full text - search for articles related to antibacterial prevalence,
antibacterial soap, and antibiotic resistance
○ Cite the source
○ Created annotation for each source, explaining how that source is relevant

Observations:
● Initial observations- nothing visible to the human eye

Data:
● Lanyero, H., Eriksen, J., Obua, C., Stålsby Lundborg, C., Nanzigu, S., Katureebe, A.,
Kalyango, J. N., & Ocan, M. (2020). Use of antibacterials in the management of
symptoms of acute respiratory tract infections among children under five years in Gulu,
northern Uganda: Prevalence and determinants. PLoS ONE, 15(6), 1–14.
https://doi.org/10.1371/journal.pone.0235164
○ Use of antibacterials in the management of symptoms of acute respiratory tract
infections among children under five years in Gulu, northern Uganda: Prevalence
and determinants

29
 ● Yu-Ting Wang, Yu-Tzu Lin, Tsai-Wen Wan, Der-Yuan Wang, Hsu-Yang Lin, Che-Yang
Lin, Yu-Chih Chen, & Lee-Jene Teng. (2019). Distribution of antibiotic resistance genes
among Staphylococcus species isolated from ready-to-eat foods. Journal of Food &
Drug Analysis, 27(4), 841–848. https://doi.org/10.1016/j.jfda.2019.05.003
○ Distribution of antibiotic resistance genes among Staphylococcus species
isolated from ready-to-eat foods
● Kammili, N., Rani, M., Styczynski, A., latha, M., Pavuluri, P. R., Reddy, V., & Alsan, M.
(2020). Plasmid-mediated antibiotic resistance among uropathogens in primigravid
women—Hyderabad, India. PLoS ONE, 15(5), 1–12.
https://doi.org/10.1371/journal.pone.0232710
○ Plasmid-mediated antibiotic resistance among uropathogens in primigravid
women —Hyderabad, India
● Mo, S. S., Urdahl, A. M., Madslien, K., Sunde, M., Nesse, L. L., Slettemeås, J. S., &
Norström, M. (2018). What does the fox say? Monitoring antimicrobial resistance in the
environment using wild red foxes as an indicator. PLoS ONE, 13(5), 1–17.
https://doi.org/10.1371/journal.pone.0198019
○ What does the fox say? Monitoring antimicrobial resistance in the environment
using wild red foxes as an indicator
● Rameshkumar, G., Ramakrishnan, R., Shivkumar, C., Meenakshi, R., Anitha, V.,
Venugopal Reddy, Y. C., & Maneksha, V. (2016). Prevalence and antibacterial
resistance patterns of extended-spectrum beta-lactamase producing Gram-negative
bacteria isolated from ocular infections. Indian Journal of Ophthalmology, 64(4), 303–
311. https://doi.org/10.4103/0301-4738.182943
○ Prevalence and antibacterial resistance patterns of extended-spectrum
beta-lactamase producing Gram-negative bacteria isolated from ocular infections

Conclusions / Summary:
● I gained a greater understanding for how to graph using excel and I learned how to
properly create an annotated bibliography.

30
Laboratory 9 Isolation of DNA
Purpose of Lab / Objectives / Hypothesis:
● Isolate DNA from your cheek cells
● Explain the process, and list the steps, of DNA isolation

Pre-Lab Lecture Notes:

● Biotechnology- the manipulation of organisms to do practical things and to provide useful
products
● Genetic engineering- the direct manipulation of genes for practical purposes
● Molecular biology- the study of molecules critical to life, at the heart of genetic
engineering
● DNA= deoxyribonucleic acid: molecule of heredity
● discovery=1953, Rosalind Franklin, Watson and Crick
● Biomolecule- nucleotides:
○ Phosphate group
○ Sugar
○ Nitrogen containing base
○ Different nucleotides
● DNA is transcribed into rRNA (ribosomal), mRNA (messenger), and tRNA (transfer)
● RNA is then translated into protein
● Proteins: contractile, transport, structural, protective, and regulatory (if needed see
biomolecule section in book)
● Paleontology, evolution, medical science conservation biology, species interactions
(contributions of genomics and proteomics to biology)
● Steps to DNA extraction: cell harvesting, cell lysis (break cells open), DNA precipitation
○ Lysis buffer = soap
● Extract your own DNA:
○ Swish with salt water for 1 minute
○ Spit into beaker
○ Add swirl of soap (no bubbles)
○ Add alcohol on top of saltwater - wait 2.5 minutes
○ White clumps = DNA
● DNA visualization- have gel matrix, put DNA sample into well, and by means of electrical
current it moves DNA through the gel, DNA is negatively charged so will be attracted to
the positive side
● See slide 15 for gel electrophoresis
● The bigger you are, the slower the fragments are able to move
● Determine size of DNA pieces by running it through the gel
● Slide 16 shows relationship between DNA fragment size and distance

31
 ● Cold temperature inhibits DNA breakage (stabilizes DNA)
● Sodium ions neutralize the negative charge of DNA to facilitate precipitation- helps DNA
come out of solution
● Detergent emulsifies cell and nuclear membranes of cells- detergent disrupts
phospholipid bilayers
● Purpose of meat tenderizer is to digest associated proteins that bind the DNA
● Purpose of ice cold 95% ethanol is to precipitate the DNA from the filtrate
● Correct way to spool the DNA- slowly, using a constant motion and pressure to collect
the precipitate
● Purpose of installing comb- to create a well or pockets in the gel where samples can
later be placed
● Ethidium bromide is needed to see the DNA bands in the gel under UV illumination
● Gel needs to be solid and uniform, no bubbles so movement can occur
● If gel was run for too long, the sample bands would move too far and leave the bottom
of the gel
● Stop too soon and the samples won’t separate far enough to be useful, but too long and
they’ll fall off the far end of the gel leaving no useful results

Detailed Procedure:
● Extract your own DNA:
○ Swish with salt water for 1 minute
○ Spit into beaker
○ Add swirl of soap (no bubbles)
○ Add alcohol on top of saltwater - wait 2.5 minutes
○ White clumps = DNA
Observations:
● Cheek cell: darker blue splotches, with even darker dot inside- nucleus

32
 ● Extracted DNA from cheek cells
Data:
● See images and notes above

Conclusions / Summary:
● I was able to isolate and extract DNA from my cheek cells, then view it through a
microscope.

Laboratory 10 Mendelian Genetics

Purpose of Lab / Objectives / Hypothesis:
● Describe simple genetic dominance, incomplete dominance, and lethal inheritance.
● Describe possible genotypes for traits inherited as dominant and recessive genes.
● Explain the importance of Mendel’s Principle of Segregation and Principle of Independent
Assortment.
● Distinguish between an organism’s phenotype and genotype.

Pre-Lab Lecture Notes:

● Mendelian Genetics- the experiments and conclusions in Mendel’s paper now form the
foundation for this
● Particulate theory- inherited characters are determined by particular factors (now called
genes), these factors occur in pairs (genes occur on maternal and paternal homologous
chromosomes), and when gametes form, these genes segregate so that only one of the
homologous pair is contained in a particular gamete
● Law of Segregation- each gamete has an equal chance of possessing either member of
a pair of homologous chromosomes

33
 ● Law of Independent Assortment- Mendel’s 2nd law, genes on nonhomologous or
different chromosomes will be distributed randomly into gametes
● Gene- unit of heredity on a chromosome
● Alleles- gene that has alternate states, contributed to an organism by its parents
● Dominant- alleles that mask expression of other alleles but are themselves expressed,
capital letter
● Recessive- alleles whose expression is masked by dominant alleles, lowercase letter
● Genotype- all the alleles present in the cell, whether they are dominant or recessive
○ Product of 2 alleles inherited from parents (if diploid)
● Phenotype- physical appearance of the trait
● Homozygous- when the paired alleles are identical
● Heterozygous- when the paired alleles are different
● First filial or F1 generation- the first generation of offspring
● Codominance- both alleles contribute to the phenotype of a heterozygote (both red and
pink show up)
○ Multiple alleles: often more than 2 alleles in a population (ex: blood types A, B,
O)
● Antigens- on the surfaces of red blood cells, determine the type of blood
● Antibodies- proteins that agglutinate certain cells (page 183)
○ Agglutination- blood clumps up, cannot survive in that situation
● Locus- location of a particular gene (ex: flower color)
● Not always possible to determine genotype from phenotype
○ Recessive allele masked by dominant allele
● Simplest case = monohybrid cross
○ A genetic cross involving a single character
● Incomplete dominance- heterozygotes have intermediate appearance; looks like a
mixture between both parents
○ Observed when the phenotype is quantitative rather than discrete
● Lethal inheritance- involves inheriting a gene that kills the offspring
● Genotype: AA, AO, BB, BO, AB, OO
● Phenotype: Type A, Type A, Type B, Type B, Type AB, Type O
● Important to be able to differentiate between male and female flies to ensure that you
have the appropriate mix of male and female flies in your cross
● The adult flies from the dihybrid cross vial were removed because the goal of the
experiment is to count the phenotypic ratio of the offspring; therefore, the parental
generation must be removed

Detailed Procedure:
● 17.2 Determine color and (shape) ratios for corn plants
○ Dihybrid-
○ Yellow/Wrinkled: 21 (21/21 = 1)
○ Yellow/Smooth: 45 (45/21 = 2.1)
○ Purple/Wrinkled: 41 (41/21 = 1.95)
○ Purple/Smooth: 143 (143/21 = 6.8)

34
 ○ Yellow-
○ Yellow/Wrinkled: 56 (56/56 = 1)
○ Yellow/Smooth: 194 (194/56 = 3.5)

○ Purple-
○ Purple/Smooth: 174 (174/76 = 2.3)
○ Yellow/Smooth: 76 (76/76 = 1)

3.
a. What is the expected genotype for the F1 generation?
RrSs
b. Will all F1 offspring have the same genotype?
Yes
c. Will all F1 offspring have the same phenotype?
Yes
4.
a. What are the predicted phenotypes for the F2 generation that is produced by
the cross RrSs X RrSs?
Red smooth, red wrinkled, white smooth, white wrinkled
b. In what ratio will they occur?
9 red smooth: 3 red wrinkled: 3 white smooth: 1 white wrinkled
5.
a. How do your data compare with those that you predicted?
My data’s ratio was 6.8: 1.95: 2.1: 1. Three parts of my ratio were
relatively close to the theoretical ratio. My ratio shows 6.8 red smooth, whereas
the theoretical ratio is 9 red smooth.
b. What are the genotypes of the F2 generation that is produced by the cross
RrSs X RrSs?
RRSS, RRSs, RrSS, RrSs, RRss, Rrss, rrSS, rrSs, rrss
c. In what ratio will they occur?
1:2:2:4:1:2:1:2:1
● 17.3 Determine blood type for ABO system
○ Patient 2: agglutinated in B (type B) more cloudy, opaque; Rh - B -
○ Patient 3: agglutinated in AB +
○ Patient 1: agglutinated in A, Rh + (agglutinated in Rh) A +
○ Patient 4: O -
○ Patient 5: A +
● Table 17.3 - Other Human Traits

Table 17.3
Phenotypes and Genotypes of Human Traits

Characteristic Your Phenotype Your Genotype

Widow’s peak No widow’s peak ww

35
Bent little finger Straight finger bb

Albinism Not albino AA/Aa

Pigmented iris Brown iris PP/Pp

Attached earlobes Unattached earlobes EE/Ee

Hitchhiker’s thumb No hitchhiker’s thumb HH/Hh

Interlacing fingers Left thumb over right thumb CC/Cc

PTC tasting Tasted it Tt

Mid Digital hair No hair mm

Dimpled chin No chin dimple mm

Six fingers 5 fingers ss

9. Which child or children could belong to a couple having AB and O blood types?
Child 1: type A and child 2: type B
10.
a. Can a person with type O blood safely donate blood to a person having type A
blood? Why or why not?
Yes, a person with type O blood can safely donate blood to a person
having type A blood. This is possible because a person with type O blood does
not have A or B antigens in their blood. This person does have antibodies against
both A and B antigens.
b. Which blood type would be a universal donor?
Type O-negative because this type of blood does not have any antigens.
c. Which blood type would be a universal recipient?
Type AB-positive because this type of blood does not have any
antibodies that will attack other types of blood.

Observations:
● For the blood typing, any blood that appeared cloudy and opaque demonstrated
agglutination of the blood cells

Data:
● Dihybrid-
● Yellow/Wrinkled: 21 (21/21 = 1)
● Yellow/Smooth: 45 (45/21 = 2.1)
● Purple/Wrinkled: 41 (41/21 = 1.95)
● Purple/Smooth: 143 (143/21 = 6.8)

● Yellow-
● Yellow/Wrinkled: 56 (56/56 = 1)

36
 ● Yellow/Smooth: 194 (194/56 = 3.5)

● Purple-
● Purple/Smooth: 174 (174/76 = 2.3)
● Yellow/Smooth: 76 (76/76 = 1)

Conclusions / Summary:
● I was able to determine phenotypic and genotypic ratios from counting corn. I did blood
typing to determine blood types based on agglutination. Finally, I was able to look at my
own traits to determine my phenotypes and genotypes of each.

Laboratory 11 Brine Shrimp Development 1

Purpose of Lab / Objectives / Hypothesis:
● Calculate and make percent salt solution.
● Calculate and make dilutions of concentrated salt solutions.
● Formulate a testable hypothesis.
● Set-up an experiment and correctly identify dependent and independent variables, and
an appropriate experimental control.
● Explain potential environmental effects on developmental processes.

Pre-Lab Lecture Notes:

● M1V1 = M2V2
● Brine shrimp are typically found in brackish water
● Non Ideal environmental conditions - high salinity or food shortage could lead to death
● Umbrella stage- hatching membrane remains attached to the cyst
● Nauplius- young brine shrimp
● Testing how various levels of salinity affect development and hatching of brine shrimp
eggs

Detailed Procedure:
● Create salt solutions and dilutions based on calculations
● Label 4 beakers with initials and type of treatment
● Put each solution into their respective beaker
● Use the refractometer to measure the salinity of each salt solution and record in the data
sheet
● Use the pH strips to measure the pH of each solution and record in the data sheet
● Add ½ scoop of eggs to each beaker and carefully stir
● Transfer 1 mL of each solution with eggs, using pipette, to petri dish and view under
dissecting microscope
● Count eggs and shrimp, record into data sheet
● Return the shrimp and solution to the beaker
● Cover the beakers loosely with plastic wrap
● Return to count the eggs in 48 hours and 72 hours and repeat the same process

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Observations:
● Eggs that appeared to be hatching but were still attached to the cyst looked like
umbrellas
● Nauplius shrimp were rapidly moving around

Data:
●

Conclusions / Summary:
● I was able to use the equation M1V1 = M2V2 to determine and create dilutions of
concentrated salt solutions. I started with 20% and made solutions of 10%, 5%, and 1%.
I properly used a dissecting microscope to count eggs, umbrella shrimp, and nauplii
shrimp. This data can now be analyzed to see relationships between percentage of salt
solution and development of brine shrimp. Brine shrimp prefer a naturally salty
environment. If the environment is too salty or not salty enough, they do not survive and
develop as well.

Laboratory 12 Brine Shrimp Development 2

Purpose of Lab / Objectives / Hypothesis:
● Identify the components of a lab report
● Describe the information that goes into each section of a lab report
● Properly use in-text citations

Pre-Lab Lecture Notes:

● Components of lab report:
○ Introduction- what is the problem?
○ Methods- when, where and how?
○ Results- what did you find out?

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 ○ Discussion- what does it all mean?
○ Literature cited- CSE citation style
● Figure should not have bright colors, include legend

Detailed Procedure:
● Rinse brine shrimp down the drain
● Wash beakers
● Dispose of petri dishes and plastic wrap in the garbage can

Observations:
● N/A

Data:
● N/A

Conclusions / Summary:
● I was able to create a formal lab report that includes all of the proper components.

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