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All AP Biology
Lab
Write-Ups
Years 2022-2023

By: Paige Hinckley

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Index:
- Cell Size: An Approach to Understanding a Cell’s Limitations: Pg 3-7

- The effects of Catalase on the Decomposition of Hydrogen Peroxide: Pg

8-21

- Lights Effect on Photosynthesis using leaf disks: Pg 22-29

- Cellular Respiration: Pg 30-36

- Bacterial Transformation Using of E Coli: Pg 37-42

- NaCl Effect on the growth and Development of Brine Shrimp: Pg 43-49

- A Common Ancestry Between Organisms: Pg 50-56

- Isopod Reactions to Different Environments which includes the experimentation

between a Wet vs. Dry environment as well as an Acidic vs. Basic Environment: Pg

57-66

- Using the Mark and Recapture Method in Order to Calculate the Size

of a Population: Pg 67-73
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Paige Hinckley Lab Write Up #1 10/5/2022

Partner: Mikayla Lawrence

Cell Size: An Approach to Understanding a Cell’s Limitations

Abstract: During the Laboratory experiment I cut 3 different sizes of beets, ranging from 3cm to

1cm and placed them in bleach for 30 minutes. The results were to show which size of beet

would be more efficient. This would represent the reasoning as to why cells are so small, well for

efficiency. The results showed that the smaller the cell the higher surface area to ratio would be

meaning, the smaller the cell, the more efficient.

Introduction: A question was posed during this experiment, why is it that cells can not grow

infinitely in size? Well cell’s have a surface area to volume ratio that is extremely important and

keeps cells a relatively small size. The surface area allows materials to effectively diffuse

throughout the cell. It is important for small cells to have a high surface area to volume ratio for

substances to efficiently diffuse through the cell membrane. During this experiment we used

beets and placed them in bleach which would turn the red pigment in beets white. The pigment

determines the color of materials. This procedure was to test surface area in order to tell which

cell size is more effective and why cells are so small. A Null Hypothesis to this procedure is that

the surface area to volume ratio of beets to bleached beet will be no different no matter the size

of the beet. An alternative hypothesis is that the smaller the beet size the higher the surface area

to volume of beets to bleach will be.

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Materials and Procedure: In order for others to perform this procedure there are multiple

materials needed. For starters you will need fresh beets, it will work just as well with canned

beets, but fresh beets are preferred for the most accurate results. In order to cut the beet you will

need a knife, size may vary, I recommend that you use a small paring knife but any size is

perfectly fine. You will then need a ruler to measure the beet into the most accurate shapes you

can make. Next you will need bleach in order to color the beet, this experiment will work best

with a fresh bottle of bleach. Along with the bleach you will need 3 beakers and tongs. The

beakers are for holding the bleach while the tongs are for taking the bleach out of the containings

at the end of the experiment. Plastic wrap is also required to place over the 3 beakers when the

beets and bleach are in their containers so that no outside variables will affect them and taint the

results. You will also need a timer to calculate how long the beets are in the bleach. And a

calculator to calculate the surface area to volume ratio. Most importantly you will need safety

supplies including; goggles, gloves, and an apron to prevent bleach getting on your skin, clothes,

and/or eyes.

Now that you have all of your materials it's time for you to begin your experiment. First

and foremost, put on your safety gear including goggles, gloves, and an apron. After you have

done that it's time to begin. Start with cutting 3 different sizes of beets, the beets should be 3cm,

2cm, and 1cm in cube form, meaning each size should be equal and there should be a total of 6

sides per cube. One thing to note is to make sure to measure your beets after you have cut them,

you will probably get a different size and they won't be perfect but still make note of it. You

should then place each cube, there should be three, into their own separate container. Begin

pouring bleach until the cubes are completely submerged in the bleach. Then cover beakers with

plastic wrap in order to prevent outside substances tainting the results. After that, set a timer for
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30 minutes. When the timer rings and the 30 minutes is up, unwrap the plastic wrap and carefully

extract the beets from the bleach with tongs and then place the beets on a paper towel. After that

is done, take out the knife you use to cut the beet cubes and cut the beets in half, and note your

observations. Once that is done use a ruler to measure the volume of the penetrated beet cube

which will be the white part and the volume of the unpenetrated part of the beet cute which will

be the part that is still white.

Data Collection and analysis:

Cube Size Total Unpenetrated Penetrated Percent Surface SA:V

Volume Volume Volume Diffusion Area Ratio

(cm³) (cm³) (cm³) (%) (cm2)

3cm 26.97cm³ 15cm³ 11.97cm³ 44.38% 53.94cm2 2

2cm 5.7cm³ 2.7cm³ 3cm³ 52.63% 17.28cm2 3.08

1cm .64cm³ .384cm³ .256cm³ 40% 4.8cm2 7.5

How I got to that Data:

3cm

V=(3)(3.1)(2.9)=26.97 Unpenetrated V= (2.4)(2.5)(2.5)=15 Penetrated V= 26.97-15=11.97

Percent Diffusion: 11.97/26.97= 44.38% SA: (3)(3.1)(6)= 53.94 SAV: 2

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Discussion and Conclusion: After the experiment, I have found that as the cells increased in size

their surface area to volume ratio decreased. For example, the 3cm cube has a SA:V (surface area

to volume) ratio of 2, the 2cm cube has a SA:V ratio of 3.08, and the 1cm cube has a SA:V ratio

of 7.5. This happens because the more volume a cell originally has, the more difficult it is to

have substances diffuse throughout the cell. In smaller cells it is easier and quicker for materials

to diffuse in it which means it is more efficient, representing the statement that cells are small for

efficiency. But there are some errors that could have occurred during this experiment. For starters

the size of the cubes was not perfect which is an error. Leading to weird results that do not

completely support the theory as to why cells do not grow infinitely. Secondly, the saran wrap

covering the bleach and beet beakers could have had different tightness’ which could have

allowed outside elements to have an effect on the substance. Lastly, the beats could have not

been completely covered in bleach because when the bleach was added the beats floated which

can make it hard to cover the entirety of the beet.

In conclusion, these laboratory results support my hypothesis. My hypothesis was that the

smaller the beet size the higher the surface area to volume of beets to bleach will be. I do believe

my results are accurate even though the results may have flaws. But I do have plenty of

suggestions for this experiment. First of all, there should be more than one trial. If there is more

than 1 trial, the results will be far more accurate. Also the way of measuring the beet cubes could

be fixed. Instead of using a knife and rule to bindly measure the cubic lengths, widths, and

heights, we could trace a piece of paper and outline it with a piece of paper. Then on the beet you

could place the paper on the beet and cut into the beet without using a ruler to outline with a

knife. This would make the results more accurate and efficient for this experiment.
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Questions: This investigation can be continued by using other fruits like cantaloupe or

watermelon to place in bleach. This will show that this can happen in all different kinds of foods,

not just radish. It adds more variety to the experiment. We could also time the amount of time it

takes for radish to have a certain amount of penetration percentage within the cell. This could

help us conclude that different sizes in cells and a high surface area to volume ratio means higher

efficiency.

This investigation did raise a few questions. What would happen if the beets were placed in the

beaker for a longer amount of time? What would happen? Would the results be any different

from before? Also the question of, what would happen if different fruits were placed in the

bleach? Lastly, what would happen if the fruit had folds to represent other organelles like

mitochondria in the cell? This could help explain why surface area is so important.
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Paige Hinckley Lab Write-Up #2 10/31/2022

Partner: Mikayla Lawrence

The effects of Catalase on the Decomposition of Hydrogen Peroxide

Abstract: During this laboratory experiment 3 different types of procedures were performed.

During the first procedure, I dipped 5 filter papers, cut out by a hole puncher, into a potato pump

and placed them in different hydrogen peroxide concentrations that were 3%, 1.5%, 0.75%,

0.375%, and distilled water which is 0%. These results will determine the effects of substrate

concentration on the decomposition of hydrogen peroxide. Throughout the 2 procedures, I

suspended a graduated cylinder containing filtered water, dish soap, and 5 grams of potato as

well as a boiling tube containing 2mL of hydrogen peroxide in water baths that were 5℃, 20℃,

40℃, and 60℃. The results earned from this procedure will determine the effect of temperature

on the decomposition of hydrogen peroxide. Finally, during the third procedure, I set out 4

graduated cylinders with 10mL of hydrogen peroxide and gently stirred in 1-2 drops of dish

soap. Then 4 1cm potato pieces were cut up in different ways; whole, ½, ¼, and randomly sliced

into multiple tiny pieces. The purpose of the procedure is to determine the surface area’s effect

on the decomposition of hydrogen peroxide. All in all, the purpose of this experiment as a whole

was to examine the way that cells break down hydrogen peroxide by using potatoes.

Introduction: Hydrogen peroxide has multiple positive benefits which includes being an

antiseptic that prevents skin infections. However, hydrogen peroxide is a byproduct of all cells
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and can become toxic to cells. So cells need to have a way of breaking down hydrogen peroxide,

they do this by enzymes. Enzymes are macromolecules that help speed up chemical reactions by

lowering the activation energy. Catalase is the enzyme that breaks down hydrogen peroxide.

Hydrogen peroxide is a substrate which is the material that fits into catalase’s active site, the area

for substrates to bind, and breaks down into water and oxygen. However, different outside

factors affect the rate at which hydrogen peroxide is broken down, such as surface area which is

the amount of area the cell has outside allowing for more efficiency within a cell, temperature,

and substrate concentration which is the concentration of substrates within a cell, this can

increase the efficiency of enzymes. The purpose of this experiment is to look at all the

temperatures, substrate concentration, and surface area to determine how these factors affect the

rate of decomposition of hydrogen peroxide, showing how enzymes react under certain

conditions. In terms of procedure 1, a null hypothesis would be that the substrate concentration

will have no effect on the decomposition of hydrogen peroxide. I predict that if the substrate

concentration increases then the decomposition of hydrogen peroxide will increase as well. In

procedure 2 a null hypothesis is; the temperature will have no effect on the concentration of

hydrogen peroxide. I predict that if temperature increases then the decomposition of hydrogen

peroxide will increase and then decrease because enzymes have an optimal temperature. Finally,

a null hypothesis to procedure 3 is that surface area will have no effect on the decomposition of

hydrogen peroxide. But I predict that if surface area increases then the rate of decomposition will

increase as well.

#1 Materials and Procedure:

Materials: Before starting procedure one, multiple materials are needed. First and foremost, you

will need Potato extract as well as a container for the potato extract. On top of that, you will need
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to obtain an insulated container full of crushed ice and then you will place the container of potato

extract within the insulated container, preventing the enzymes from working. In addition, you

will need a plethora of different containers, including a 100mL and a 10mL graduated cylinder, 5

beakers, and five test tubes. Hydrogen peroxide will also be needed, 3% to be exact, this is the

normal percent of hydrogen peroxide available in grocery stores or local pharmacies. This

hydrogen peroxide will eventually be distributed into different dilutions between the 5 beakers.

Furthermore, filter paper will be needed because it represents how enzymes speed up reactions

so the filter paper will float to the top. To cut the filter paper you will need a hole puncher and

should punch at least 5 holes. Additionally, a paper towel will be needed to dab the filter paper

on to keep the paper from dripping the potato extract. In order to get the filter paper in and out of

the test tubes forceps or a stirring rod can be used. Finally, a stopwatch will be used to time how

long it takes for the filter paper to float to the top of the solution. You will also need distilled

water that will act as a negative control as it is known to have no effect on the filter paper.

However, before starting this experiment you need to have safety equipment. This includes

protective goggles, a lab coat, gloves, as well as the forceps that were mentioned before to not

touch the hydrogen peroxide solution.

Procedure: Now that you have obtained all the necessary materials you will need for this

procedure starts by placing the potato pulp within the container in the insulated container full of

crushed ice. Secondly, you will need to prepare a plethora of different hydrogen peroxide

solutions. First label 5 different breakers with the desired hydrogen peroxide concentration; 3%,

1.5%, .75%, .375%, and 0%. In order to make these different solutions you will start with 200mL

of 3% hydrogen peroxide. Then remove 100mL of that solution and transfer it to the 1.5%

where you would then add 100mL of distilled water and stir it. Then again take 100mL of that
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solution and place it in the 0.75% beaker then add 100mL of distilled water. Then you are going

to take 100mL out of that container, place it in the 0.375% container, stir 100mL of distilled

water and discard 100mL from that container. Finally, add 100mL of distilled water to the 0%

beaker. Now you should have a 3, 1.5, .375, and the 0 percent containers filled. Now grab your 5

test tubes, and fill them up 10mL with each solution. Then cut 5 holes out of your filter paper.

Using the forceps mentioned people dip the filter paper in the potato pump, one at a time, until

completely saturated. Then dap the filter paper on a paper towel in order to drain excess pulp, but

do not dry the paper out. Next, drop the filter paper into the 3% hydrogen peroxide, and record

the time using a stopwatch, the filter paper should rise to the top except with the distilled water,

it will never float. Do this for all solutions and then record the data within a data table.

#1 Data Collection and Analysis:

Hydrogen Peroxide Group 1 Group 2 Class Average

Dilution

0 --- --- ---

0.375 28 seconds 35.62 seconds 31.81 seconds

0.75 17.83 seconds 20.02 seconds 18.925 seconds

1.5 12.77 seconds 14.89 seconds 13.83 seconds

3 8.6 seconds 10.61 seconds 9.605 seconds

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#1 Discussion and Conclusion:

Discussion: After a close examination of this experiment. I have found that an increase in

substrate concentration leads to an increase in the decomposition rate of hydrogen peroxide. For

example, when the dilution of hydrogen peroxide was 0.375% it took the potato pulp filter paper

31.81 seconds to float to the top of the solution. Then at 0.75% it took 18.927 seconds to float to

the top, at 1.5% it took 13.83 seconds to float to the top, and finally at the top hydrogen peroxide

dilution, 3%, it only took an average of 9.605 seconds to float to the top the hydrogen peroxide

solution. This occurs because when hydrogen peroxide, a substrate, comes into contact with

catalase, an enzyme, the enzyme breaks down the hydrogen peroxide allowing the filter paper to

float. If the concentration of substrates increases then it is easier for the catalase to take the

substrate into its active site and speed up the reaction, which is why the decomposition of

hydrogen peroxide will increase as the substrate concentration is increased. Of course, like any
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experiment, there could have been some errors. One error that could have been made was that the

hydrogen peroxide concentrations were not accurate which could affect the rate at which the

filter paper floated to the top of the hydrogen peroxide. Another error that could have been made

is that the potato pulp could have been warmer and the ice bath could have melted. Making the

enzyme start to react on its own could produce no results for this procedure. Lastly, the filter

paper could have been dried too much on the paper towel and most of the enzyme may have been

absorbed making the reactions take a lot longer than necessary.

Conclusion: In conclusion, the results of this experiment support my hypothesis. I hypothesized

that an increase in the substrate concentration would lead to an increase in the decomposition of

hydrogen peroxide. I believe that the results I have obtained are accurate and I do not believe

there were any significant mistakes within the experiment itself. I do have a few suggestions for

the experiment itself though. For starters, there should be at least three trials per dilution of

hydrogen peroxide, this will lead to more accurate results and allow room for mistakes to be

noticed if any mistakes are made. Secondly, the potato pulp should be stored in a place where

there is no fluctuation between temperatures. Yes, an ice bath does keep the pulp cold but ice can

eventually melt, causing the enzymes to start working. Instead, the potato pump should be stored

in a fridge where the temperature can be exactly controlled. This would lead to less room for

error. Overall, this experiment is very well done.

#2 Materials and Procedure:

Materials: Before beginning this procedure you will need a plethora of supplies. For starters,

you will need a potato, the potato should be fresh. In order to chop the potato you will need a

knife and chopping board, this should allow you to cut up the 5 grams of potato needed for this

experiment. Next, you will need 3% hydrogen peroxide, you should have this from the first
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procedure, as well as filtered water. Additionally, you will need dish soap, it doesn’t matter what

brand it is. But a pipet should be used to extract the dish soap, and be disposed of after the

experiment. In addition, 4 graduated cylinders and well as 4 boiling-proof test tubes with and

stand to hold them in place will be needed. The water baths are also important, you will need 4

different temperatures, 5℃, 20℃, 40℃, and 60℃. Finally, A balance is also needed as well as

weight boats that should be placed on top of the balance, this should allow you to measure the

initial and final volumes of the beaker. However, don’t forget to wear your safety goggles, lab

coat, and gloves.

Procedure: If you haven’t already set up the four water baths mentioned before with the

temperatures, 5℃, 20℃, 40℃, and 60℃. After doing that add 20mL of filtered water to a

graduated cylinder, then add dish soap to the graduated cylinder. Next, using a knife and a

chopping board, finely slice a potato and weigh out five grams of the potato using a weight boat

placed onto a balance, set this aside for later. In a boiling tube, you will need to add 2mL of 3%

hydrogen peroxide. Now you will add the 5 grams of chopped potato and add it in the graduated

cylinder that already contains both the dish soap and the filtered water. Then, suspend both the

boiling tube as well as the graduated cylinder in the water bath. I recommend that you start at

5℃ and work your way up in temperature. Suspend these solutions for approximately 3 minutes

or until they reach the desired temperature. Once the solutions reach the desired temperature,

immediately read the volume of the solution, then place both the boiling tube and the graduated

cylinder back into the water bath and set a timer for exactly 2 minutes. After the two minutes are

up take both of the containers holding the solutions and record the volume. Finally, subtract the

initial values from the final values to determine the amount of foam generated, and record the
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volume in cm^3. But remember you need to do this for all of the temperatures; 5℃, 20℃, 40℃,

and 60℃.

#2 Data Collection and Analysis:

Temperature (°C) Initial Volume (cm^3) Final Volume (cm^3) (Final-Initial)

5 20.1 22.2 2.1

20 20.2 36.4 16.2

40 20.0 30.1 10.1

60 20.1 21.0 0.9

#2 Discussion and Conclusion:

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Discussion: During this experiment, I have observed the effects of temperature on the

decomposition of hydrogen peroxide. I have found that an increase in temperature will lead to an

increase in the decomposition of hydrogen peroxide and then a decrease in hydrogen peroxide,

this is due to an enzyme's optimal temperature. For example, at a temperature of 5℃ 2.1mL of

foam was generated, at 20℃ 16.2mL of foam was produced which is a massive difference

compared to 5℃. After that, the foam amount decreases, at 40℃ 10.1mL of foam is produced,

and at 60℃ only .9mL of foam is produced. The reason behind this is that all enzymes have an

optimal temperature, in which they function the best. However, if that temperature becomes too

great for an enzyme then the enzyme can start to denature, meaning its amino acid strands begin

to unravel, leaving the enzyme functionless. In this case, the optimal temperature is 20°C due to

the production of foam representing the decomposition of hydrogen peroxide. However, this

experiment has many flaws that need to be addressed. For example, maintaining the temperature

of water baths can be extremely difficult especially when you only have a stovetop, the water

bath wouldn’t stay at a constant temperature, changing the results completely. Additionally, most

of the time the potato weight will not be the same for every single water bath, this will create

fluctuation between results making the results unusable and insignificant.

Conclusion: In conclusion, these results have supported my hypothesis. I predicted that if

temperature increases then the decomposition of hydrogen peroxide will increase and then

decrease because enzymes have an optimal temperature. Also, I believe that the results I have

retained are accurate but there may be some minor mistakes within this experiment itself. I have

multiple ideas for improving this experiment. First of all, the temperature of the water should be

easier to heat up. For example, there should be something that keeps the containers at the perfect

temperature such as an oven that maintains a certain, constant temperature because the heat of
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the water is usually fluctuating constantly without a heat source that keeps the solution at a

specific temperature. Another recommendation is that the potato should be able to measure more

accurately, most of the time starch from the potato adds a lot of weight to the potato so the potato

leaves residue on the balance fluctuating the weight. To make the weight constant the outside of

the potato should be dabbed with a paper towel so not a lot of residue is left on the scale and the

weight can remain constant. Last but not least, there could always be more trials which would

lower the window for errors, making the experiment more plausible.

#3 Materials and Procedure:

Materials: Compared to procedures 1 and 2 procedure 3 doesn’t require many materials. First

and foremost, a potato is needed as well as a knife and a chopping board to cut up the potato.

You will also need 3% hydrogen peroxide and dish soap. For the dish soap, a pipet is needed for

precise drops of dish soap but after the pipet is used you are able to dispose of it because the

pipette does not clean out easily. You will need 4 graduated cylinders. Finally, for safety

purposes, don’t forget to wear safety goggles, a lab coat, and gloves

Procedure: For starters, set out the 4 graduated cylinders mentioned before. Then add 10mL of

3% hydrogen peroxide to the graduated cylinder. On top of that add 1-2 drops of dish soap to the

graduated cylinder and stir gently in order to not create any bubbles. You then want to take out

both your knife and chopping board to cut a potato. You need 4 square cubes that are 1 cm on

each side. The first cube will remain the same, the second cube will be split in half, the third

cube will be cut into fourths, and the fourth will be cut into multiple pieces. First, add the large

cube into the first graduated cylinder, labeling them might also be helpful, then set the timer for 5

minutes. 30 seconds after that add the cube that is split in half and time that for 5 minutes. 30

seconds later add the cube cut into fourths and time 5 minutes. Then 30 seconds after that add the
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potato cube that is cut up into multiple tiny pieces, and time it for 5 minutes. It is important to do

each of these cubes 30 seconds from each other so that the data doesn’t become scattered and it

makes it less stressful while recording the data. Then subtract the initial value from the final

value of the graduated cylinders to determine the amount of foam that was generated.

#3 Data Collection and Analysis:

Potato Cubes Initial Volume (cm^3) Final Volume (cm^3) (Final-Initial)

1 12.1 16.5 4.4

2 12.3 28.4 16.1

4 11.7 34.6 22.9

8 11.8 40.4 28.6

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#3 Discussion and Conclusion:

Discussion: After carefully examining this experiment I have found that when surface area

increases so doesn’t the decomposition of hydrogen peroxide. For example, when a 1cm cube

was placed into the solution of 10mL of hydrogen peroxide and dish soap the potato cube had a

foam growth of 4.4, then the 2 cubes had a growth of 16.2, a major difference from the one cube.

Next, the 4 cubes had a foam growth of 22.9, and last but not least the 8 potato cubes had a final

foam growth of 28.6. This occurrence is due to compartmentalization, which is an aspect that

allows cells to have more reactions without interfering with each other. The more surface area

allows the catalase enzyme to interact with the hydrogen peroxide substrate allowing for a higher

amount of hydrogen peroxide breakdown when the surface area increases. There are very few

issues that could have arrived during this experiment. For starters, the most common issue is the
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size of the potato cubes. Most of the potato cubes will not exactly be 1cm before being cut up.

Additionally, even when the cubes are cut up the sizes will not be the same for each cube. This

will create wacky results that cannot be used to support or refute my hypothesis. Lastly, the dish

soap could be stirred to the point where it has bubbles making this experiment ineffective, but it

is an easy fix and a new solution of hydrogen peroxide and dish soap can be made.

Conclusion: In conclusion, the results of this experiment support my hypothesis. I predicted that

if surface area increases then the rate of decomposition will increase as well. I know that my

results are accurate as well because I measured every piece of 1cm potato thrice. In terms of

improvement, I do not have much to say. If I were to criticize one aspect of this experiment it

would be the measuring of the potato cubes. Instead of measuring a potato with just a ruler and a

knife, there should be paper stencils to draw in order to trace the shape of the potato; or even

better, a cookie cutter that can make the perfect shape without any errors. And of course, this

experiment could always have had more trials for fewer errors and more significant information

in research terms.

Questions:

This investigation can be continued by testing different factors that will affect the rate of

enzymes. For example, pH would be an experiment and is exactly like temperature in the way it

impacts enzyme activity. They both have an optimal number at which the enzyme performs best.

It would also be interesting to try this experiment with an inhibitor. In order to show the effect of

an inhibitor on an enzyme. Also in procedure 2, it would’ve been interesting to increase the

temperature beyond 60℃ to see the effects of catalase fully denaturing.

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What questions did this investigation raise: During this investigation, a few questions were

raised. What would happen if we used a different enzyme such as lactase instead of catalase? Or

what if we tried to experiment with lactase and its effects on breaking down lactose since a lot of

people in this world have lactose intolerance? Lastly, instead of potatoes what would happen if

we used watermelon instead?

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Paige Hinckley Lab write-up #3 11/07/2022

Partner: Mikayla Lawrence

Lights Effect on Photosynthesis using leaf disks

Abstract: During this laboratory experiment I cut 20 small holes out of a spinach leaf, using a

hole puncher. The leaves were then placed in a sodium bicarbonate-water solution and separated

into 2 beakers filled to 150mL. 10 of the 20 spinach leaf disks were placed into one beaker and

the rest were placed into the other breaker. One of the beakers was left in the sun for 15 minutes

while the other was in complete darkness. These results would represent the importance of light

during photosynthesis. The results showed that photosynthesis cannot happen without light.

Introduction: A question posed within this experiment was; what is the relationship between

light and photosynthesis? Well, light is extremely important for photosynthesis to be able to

occur. Photosynthesis is the process in which plants and some prokaryotes produce their own

food in the form of glucose, essential to all life. It does this in two phases, the light reaction and

the Calvin cycle. The Light reactions occur in the thylakoid membrane within a chloroplast. This

reaction takes in CO2, solar energy, and water to create the products of ATP and NADPH which

will then go on to help the Calvin cycle, taking place within the stroma in a chloroplast. The

Calvin cycle will use ATP and NADPH to produce G3P which is needed for the formation of

glucose, but that's not all. The Calvin cycle will also produce ADP and NADP+ needed for the

light reactions. These reactions cannot occur without each other. However, during this

experiment, we are focussing on the Light reactions more specifically. During this experiment, I

used spinach leaves in the sodium bicarbonate water solution to test the effect of solar energy on

photosynthesis. During Photosynthesis oxygen is produced as one of the products so the leaf
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disks that are able to photosynthesize will float to the top. A Null hypothesis to this experiment is

that light will have no effect on the occurrence of photosynthesis. While I predict that an increase

in light exposure will increase the occurrence of photosynthesis.

Materials and Procedure:

Materials: Before starting this experiment you will need some supplies. For starters, you will

need spinach leaves, it doesn't matter if they are in a bag or not. However, you will need a hole

puncher in order to cut out small leaf disks. Next, you will need sodium bicarbonate or baking

soda, many people have this at home but it can be bought in your local grocery store. Another

household item you will need is dish soap, the brand doesn’t matter. Additionally, you will need

things that do not have an expiration date and can be found in your house or in a school lab. A

scale and weighing paper will be needed first to measure 1.5g of sodium bicarbonate. In order to

hold a solution you will need two different types of beakers, one able to hold 500mL of solution,

this is where the main solution will be made, and two beakers that can hold 200mL of solution,

which will hold the leaf disks and solution. In order to stir the solution you can use a spoon or

stirring rod. A pipette will also be used to place two drops of dish soap into the solution, the

pipette can be disposed of after use. Now to extract the oxygen from leaf disks a plastic syringe

will be needed, the syringe shouldn’t have a needle. Remember that light is the most important

aspect of this experiment. If it is a nice and clear day outside, bring one of the solutions outside

while leaving the other one in complete darkness, like in a cabinet. However, if it is not nice

outside, you can use an ordinary lamp. Lastly, safety is the most important. Make sure to wear

goggles, a lab coat, gloves, and use a paper towel if anything spills.
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Procedure: Now that you have collected all the materials needed for the experiment. First of all,

take 3-4 spinach leaves and punch 20 holes in them using a hole puncher, making sure to punch

between the veins. Next, you need to prepare a sodium bicarbonate and water solution. To make

this, weigh 1.5g of sodium bicarbonate on a scale and add it into the 500mL beaker. Then pour

300mL of water into the beaker containing the sodium bicarbonate, Stir gently until completely

mixed. Next, add 2 drops of dish soap into the solution and stir it carefully to avoid bubbles.

Now split the mixture into 150mL each within the two 200mL beakers. If there are bubbles

alternate between the large beaker and the small beaker to filter the bubbles out. In order to

extract oxygen from the leaf disks find the string and remove the plunger. Then fill the syringe

with the 20 disks, but I have found it easier to separate the disks into ten first and don’t do all the

disks at once. Next, add the sodium bicarbonate and water solution into the syringe about ⅓ of

the way full, and put the plunger back in. Hold the syringe upward and gently expel the

remaining air from the tip of the syringe. Then place your finger on top of the syringe and pull

the plunger back, creating a vacuum, hold it for 20-30 seconds. The disks should sink if they

don’t try it a second time but if it doesn't work after that hole punch 20 new leaf disks. Now

remove the plunger and gently separate the solution from the disks. Put the solution back into the

beaker and separate disks into groups of ten in each 200mL beaker. Finally, place one beaker in

light and watch the disks float up for 15 minutes. Then place the other disks in a dark area with

no light. Record your data.

Data Collection and Analysis:

 Hinckley 25

Time (min) # of leaf disks # of leaf disks

floating floating (Dark)

(Light)

1 0 0

2 1 0

3 6 0

4 9 0

5 9 0

6 9 0
 Hinckley 26

7 9 0

8 9 0

9 9 0

10 9 0

11 9 0

12 9 0

13 9 0

14 9 0

15 9 0

Discussion and Conclusion:

After this experiment, I have found that in order for photosynthesis to happen light is required.

Solar Energy allows for photosynthesis to occur. For example, I have found that 9 out of the 10

disks in light float to the top telling me that they went through photosynthesis since one of the

products of photosynthesis is oxygen and oxygen makes things float. Also in the beaker where

there was no light photosynthesis did not even occur because after 15 minutes no leaf disks were

floating, unlike the disks in light. Encapsulating the idea that photosynthesis requires light in

order to make glucose (C6H12O6) and oxygen (O2). I believe that my results are accurate,

however there are still plenty of errors that could have occurred. First of all I could have not
 Hinckley 27

extracted all of the oxygen out of the leaf disks. This would cause all of the leaf disks to float

even the ones in the dark, which would then taint all of the data collected. Also some light may

have been able to go to the beaker that was supposed to stay in the dark, this could cause the

leaves to have absorbed light leading them to perform photosynthesis making the experiment

inconclusive. Lastly, I could have punched a hole through the veins of the leaf causing the leaf to

have other liquid substances mixed in within the disks, this would then taint the data collected

from this experiment.

In conclusion, these laboratory results support my hypothesis. I predicted that in the presence of

light the leaf disks would be able to perform photosynthesis. For example, as time increased the

number of leaf disks floating increased for the breaker in light but stayed the same for the beaker

in the dark. Before discussing improvements that could have been made to this experiment I

would like to discuss questions posed by this experiment. For starters, the independent variable

was the light while the dependent variable was the flotation of disks representing the occurrence

of photosynthesis. Secondly, dish soap was added to the solution to stimulate photosynthesis by

coating the leaf with a thin layer of soap which will then help sodium bicarbonate stick to the

plant disks. Oxygen is a variable in light reactions, in this experiment oxygen is a product that

comes from water splitting, a process that adds a concentration gradient to light reactions. So the

purpose of creating a vacuum seal was to extract all oxygen from the plant allowing the plant to

sink. The plant sinks because oxygen is a gas that makes things float and in this case when

oxygen isn’t present the leaf disks wouldn’t float. Lastly, leaf disks in the light floated because

they were able to absorb light energy and with the help of both light energy, CO2, and water the

plants were able to perform photosynthesis using both the light reaction and Calvin cycle.

However, the plant disks that were in the dark didn’t folate because they weren’t able to absorb
 Hinckley 28

light, preventing them from going through photosynthesis. Now how can this experiment be

improved? For starters, this experiment should have been repeated at least two more times for

more accurate results. Additionally, the leaves could have been put into a vacuum chamber to get

rid of all oxygen; this would make the probability of error decrease. Lastly, the spinach leaf disks

could have been kept in their specific light requirement for a longer period of time, this would

completely prove that photosynthesis cannot happen without the presence of light.

Questions: There are numerous ways this investigation can be continued. An example of this is

using different types of leaves. For example, maple leaves, basil leaves, or flower leaves. The

effects of pH on photosynthesis would also be a viable investigation. More importantly, different

colors of light at different intensities would be something interesting to experiment with. For

example we could use green, violet, blue, and orange light to see what color plants like more in

order to perform photosynthesis.

This investigation raised a few questions. First and foremost, what would happen if the leaf disks

were placed into different colors? What would happen if we experimented using a whole leaf

instead of leaf disks? What would happen if we used leaves that were different colors? Would

there be a difference? All of these questions could give us a better understanding of

photosynthesis and the factors that affect it.

Works Cited

“Baking Soda, a Home Remedy Fungicide - the Cornell Formula Baking Soda, a Home Remedy

Fungicide - the Cornell Formula.” Garden Myths,

 Hinckley 29

https://www.gardenmyths.com/baking-soda-home-remedy-fungicide-cornell-formula/.

Accessed 7 November 2022.

 Hinckley 30

Paige Hinckley Lab Write-Up #4 11/16/2022

Partner: Mikayla Lawrence

Cellular Respiration

Abstract: During this laboratory experiment, I placed different substances into three vials

containing steel washers. The first one contained glass beads, the second, germinating seeds, and

the third was a mixture between beads and non-germinating seeds. Each of these substances was

then surrounded by cotton mixed with potassium hydroxide on the bottom and non-absorbent

cotton on the top. The vial then had a pipette with a stopper on the top, sealing the substance

within the vial. Then place all three vials in a bin full of water. This experiment will determine if

germination has an effect on the rate of respiration.

Introduction: A question was posed during this experiment; does germination affect the rate of

respiration? Cellular respiration is a process in which organisms break down glucose in order to

produce ATP, adenosine triphosphate. This will then fuel the performance and functions within

the cell. ATP is also needed for photosynthesis and all the functions requiring energy without it

an organism would not be able to survive. However, cellular respiration isn’t just a singular

phase; it contains a total of three phases; Glycolysis, Citric Acid Cycle, and Oxidative

phosphorylation. Glycolysis is the first process and breaks down glucose into 2 pyruvates. An

intermediate reaction is pyruvate oxidation which then breaks down the pyruvate into acetyl CoA

which is needed by the Citric Acid cycle, also known as the kreb cycle. The citric acid cycle uses

the Acetyl CoA to produce 2ATP, CO2, and the two electron carriers, NADH and FADH2. These

carriers will help the final process of cellular respiration. The final step in cellular respiration is

oxidative phosphorylation. The process uses H+ electrons carried by the carrier molecules,
 Hinckley 31

NADH and FADH2. This process will produce the highest amount of ATP ranging from 26-28

ATP. During this experiment the aspect of cellular respiration is being tested within plants. The

CO2 produced during cellular respiration will represent the rate of respiration. A Null hypothesis

is that germinating and non-germinating peas have no effect on the rate of cellular respiration. I

predict that the germinating peas will have a higher rate of cellular respiration than the non

germinating peas.

Materials and Procedure:

Materials: Before performing this experiment you will need a plethora of materials. For starters,

you will need 3 separate vials along with three metal washers, the metal washers should be

tapped onto the bottom, this will prevent the vials from floating up later on in this experiment.

Along with the vials and washers you will need a stopper as well as a pipette. In order to place

these two things together you can use simple waterproof tape. With both the metal washer vials

and the stoppers you are creating a respirometer. Now you will need the substances going into

the respirometer. Beads are going to be a control, the beads can be glass or plastic. Additionally,

Non Germinating peas are needed, you should place more than 20 of the non germinating peas in

the water to make them germinate because you will need 20 germinating seeds for this

experiment, but make sure you have plenty of non-germinating ones leftover. Next you will need

a 100mL graduated cylinder, this is where the substances will be contained. Surrounding the

substances you will need absorbent cotton below the substances and non absorbent cotton above

the absorbent cotton so sodium hydroxide doesn’t interfere with the substances. As mentioned

before sodium hydroxide is needed, this is a highly toxic substance so it requires a laboratory
 Hinckley 32

pipette. However, sodium hydroxide can be bought on amazon. In addition, a large bucket for

water will be needed as well as a thermometer to test the temperature of the water. The

thermometer should stay in the water throughout the entire experiment. This will check to see if

the water temperature is constant. Lastly, for safety a laboratory jacket, gloves, goggles, and

paper towels are recommended.

Procedure: Now that you have all of the material needed for this experiment it is time to begin.

For starters, we are going to begin by measuring the amount of substances for each. Fill a 100mL

graduated cylinder with 50 mL of water then place 20 germinating peas in the graduated cylinder

and measure how high the water goes when you add the germinating peas. Then take the peas out

and place them on a paper towel. Now refill the graduated cylinder with 50mL of water and

place the amount of beads needed in order to increase the water levels to the same height of the

germinating peas and place them on a paper towel. Repeat the process with beads but with the

non-germinating peas and the beads combined. Fill the graduated cylinder until it reaches the

same level as germinating peas. You do not need the same amount of non-germinating seeds and

beads though. After this is completed, fill each vial with a layer of absorbent cotton, then grab a

pipette and add .5mL of sodium hydroxide to each vial. Next, add a layer of non-absorbent

cotton on top, this will prevent sodium hydroxide from interacting with the substances. Finally,

place the stoppers connected to the pipettes onto and seal the vials, making sure that the stoppers

do not pop off. Now you have a respirometer. Set the respirometer inside the bucket of water that

should have a thermometer in it making sure that the water temperature is constant. Place the

respirometers on their sides for 5 minutes. After that time is done, submerge the respirometers

completely underwater, due to the five minutes the respirometers spent tilting a bubble should
 Hinckley 33

form within the pipette. Even five minutes for 20 minutes observe the bubble and write down

how far the bubble has gone. Record these observations in a table.

Data Collection and Analysis:

Temp Time Beads Beads Germinating Germinating Germina Beads N Beads Beads N

Alone Alone Peas Peas ting Peas Dry Peas Dry Peas Dry Peas

20 x Reading at Difference Reading at Difference Correcte Reading Difference Correct

time time d at time Difference

Differen

ce

20 0 8.85 x 8.75 x x 8.0 x x

20 5 8.8 0.05 7.25 1.5 1.45 7.8 0.2 0.15

20 10 8.75 0.10 6 2.75 2.65 6.9 1.3 1.2

20 15 8.7 0.15 4.75 4.0 3.85 5.9 2.1 1.95

20 20 8.3 0.55 3.75 5.0 4.45 5.1 2.9 2.35

Discussion and Conclusion:

Discussion: After close examination, I have found that germination seeds perform cellular

respiration at a higher rate than the non-germinating seeds. For example, during the 20 minute

interval the bubble within the pipette traveled down 5mL, with a correct difference of 4.45. The
 Hinckley 34

bubble went from 8.75mL on the pipette to 3.75mL on the pipette. This is due to the O2

consumed during this experiment. In order for cellular respiration to produce the maximi=um

amount of ATP in germinating seeds, the seeds need to intake oxygen. So when the amount of

oxygen decreases the bubble will go down the pipette. Showing that the germinating seeds

performed cellular respiration more efficiently than the non-germinating seeds. The

non-germinating seeds had the bubble in the pipette move down 2.9mL from where the bubble

first started at, 8mL. The corrected difference is 2.35. Overall the germinating seeds had a

decrease of 4.45mL while the non-germinating and beads had a decrease of 2.35mL. I am not

completely sure if my results are accurate so there's a plethora of errors that could have occurred.

First and foremost, the stoppers on the vial could have come undone, this happened for my

germinating seeds so I needed to borrow data from another group. Overall, if the stopper doesn’t

stay on the result of that substance can become extremely tainted. Secondly, I could have used

absorbent cotton instead of non absorbent cotton to place on top of the sodium hydroxide cotton.

This could cause all of the vials to have inefficient data, making this experiment useless. Lastly,

the temperature of the water could fluctuate and a spike in temperature could cause the bubble in

the pipette to speed up or slow down, making all data strange and disordered.

Conclusion: In conclusion, these laboratory results support my hypothesis. I predicted that the

germinating peas will have a higher rate of cellular respiration than the non germinating peas.

For example, as the time increased during this experiment the bubble in the germinating seed

respirator decreased greatly, more than the non-germinating seed respirator. Before discussing

improvements that need to be done to this experiment, I would like to discuss the questions that

were posed by this experiment. First of all, germination does have an effect on respiration. This

is due to the seeds needed to grow into a plant. During germination plants are not able to perform
 Hinckley 35

photosynthesis yet, which allows them to make glucose, however with stored energy within the

seed the seed is able to bring that down into ATP through the process of cellular respiration.

Additionally, the statement, “Dormant seeds do not respire” is incorrect. Due to the results the

dormant seeds still take in oxygen in order to perform respiration even though non-germinating

seeds do not go through as much cellular respiration as germinating seeds do. Secondly, sodium

hydroxide was added to the respirometers because it absorbs carbon dioxide. This will then

change the volume of gas within the respirometer, allowing the volume of gas to be directly

related to the amount of oxygen consumed. Now, if the temperature of the water increases within

the respirators, I predict that the rate of respiration will increase as well up until a certain point. If

it gets two hot then the proteins that help transfer electrons will denature and unravel which will

then change the shape and function of the protein. If the temperature of the water decreases, I

predict that the rate of cellular respiration will decrease because molecules will not be able to

move as quickly. Furthermore, water moved into the respirometers because as time increased

more oxygen was taken in by the seeds this led to a decrease in gas volume resulting in a

decrease of pressure as well, allowing water to move into the respirometers. Lastly, it is

necessary to correct the readings of the respirometers because the beads act as a control and if

they change that means some outside aspect is probably affecting the results so when you

subtract the bead respirometers from the other it assures that the data is accurate. Finally, there

are multiple improvements that can be made to this experiment. First of all, instead of stoppers,

cork could be used. I believe that cork would have a stronger attachment onto the vials so that

the respirometers wouldn’t fall apart. Additionally, the water bath should be contained in a more

secluded area so that air flow will not affect the temperature of the water. For example, putting

the path in a cardboard box which could prevent airflow from interfering with the temperature of
 Hinckley 36

the water. Of course, this experiment could have at least 2 more rounds of this experiment for

more accurate results.

Questions:

This experiment can be continued by changing certain factors within this experiment. For

starters, instead of using germinating pea seeds we could use watermelon seeds, pepper seeds, or

strawberry seeds. Would the rate of cellular respiration be different even if the size of seeds are

smaller? Also, instead of using beads, marbles could be used instead. The respirators could also

be derived from light. This could show that germinating seeds are performing cellular

respiration, supporting the idea that plants can perform both photosynthesis and cellular

respiration.

What questions did this investigation raise: During this investigation a few questions were

raised. What would happen if different seeds were used? What if sodium hydroxide wasn’t used?

How different would the results be? What would happen if the substances were the only things

within the respirators and not the cotton?

 Hinckley 37

Paige Hinckley Lab write-up #5 2/5/2023

Bacterial Transformation Using of E Coli

Abstract: During this laboratory experiment, I used four different plates with each one full of. E

Coli bacteria. The only difference is the substances being used. The first plate only contained LB

which is the nutrient broth. The second one contained both the LB of the previous plate and

AMP, which is a substance that prevents cell growth. Therefore the second plate will most likely

be empty. The third plate is the same as the second, however, this plate contains bacteria that are

thriving; this is due to the addition of the pGLO plasmid in the plate. Then the fourth plate

contains LB, AMP, and ARA. The ARA is a substance used in jellyfish to make them glow under

UV light. These substances are then placed under a UV light for observation. Overall, this

experiment will allow me to look at the bacterial transformation within E Coli.

Introduction: A question was posed during this experiment; why did the -DNA bacteria not

survive while the +DNA bacteria did? Well, this is all due to transformation. Transformation is a

process in which prokaryotic cells can exchange genetic material through horizontal gene

transfer which is a process where a gene or genes from one organism is transferred to another

organism. In the case of transformation, when a bacteria sheds a plasmid another plasmid can

uptake the shredded plasmid from its environment. This is called transformation.Transformation

is an important aspect for bacteria because it provides the bacteria with the ability to survive

tough conditions and become stronger and more modified. However, during this experiment,

multiple different plates containing bacteria were tested. However, the one containing LB was

not able to survive and this was due to AMP, which prevented the growth of the bacteria.
 Hinckley 38

However, the other plate that contained both LB and AMP flourished. This was due to a

successful horizontal gene transfer in which the LB/AMP bacteria plate was able to inherit the

pGLO plasmid, giving it the ability to survive and flourish. With that being said the exchange of

genetic material between the bacteria allows bacteria to survive. The Null hypothesis would be

that there is no difference between the Bacteria containing the pGLO plasmid and the bacteria

that doesn’t contain the pGLO plasmid. I predict that the bacteria that contains the pGLO

plasmid will be able to produce bacteria that contains a resistance to AMP compared to the

bacteria without the pGLO plasmid.

Materials and Procedure:

Materials: Before beginning this experiment you will need to gather a plethora of materials.

First and foremost, you will need E coli starter plates, these plates will most likely be prepared

for you and will already contain a growing bacteria culture of E coli. Next, you will need 4 agar

plates. 2 of these plates should just contain LB and the other 2 plates should contain a mixture of

both LB and AMP. The AMP symbolizes ampicillin, which is supposed to stop the growth of

bacteria. Additionally, you should have a CaCl2 (Calcium Chloride) solution which will help

open up the plasma membrane in order for the bacteria to absorb a new plasmid. Also, it is

important to note that this should be kept on ice so the CaCl2 doesn’t start working yet. Then

you will need an LB nutrient broth, a solution that can be used for bacterial growth. Now you

will need a sterile inoculation loop which is used to spread the E Coli culture onto the plates, but

be sure to do now use the inoculation loop more than once and change the loop whenever you're

working with a different plate. Next, a pipet is needed to transport substances throughout this

experiment. Now, in terms of a water bath, you will need microcentrifuge tubes and a floating
 Hinckley 39

holder for the water bath; these will contain the -DNA and +DNA parts. Additionally, the water

path will need to have hot water that is 42 degrees celsius, this temperature can be monitored

using a thermometer. Now, you will need ice which will be used to shock the microcentrifuge

tubes. Of course, you will also need the pGLO plasmid which contains AMP resistance, this is an

essential part of this experiment. A new incubator will be needed that has the ability to be 37

degrees Celsius. An incubator can usually be found by asking a science lab or even a farmer.

Lastly, you will just need bleach, this can be found in your local grocery store. However, don’t

forget safety. You must wear goggles, a lab coat, gloves, and if you have long hair you need a

hair tie. In order to get rid of the material once the experiment is completed use Hazard Waste

Disposable bags.

Procedure: Once you have collected all of your materials it is time to begin the experiment. First

and foremost, gather the 2 microcentrifuges and label 1 as “+ plasmid” and label the other one as

“-plasmid.” Then you should add 250ml of CaCl2 solution into each tube and then you should

play both of the tubes in a bed of ice. You will then use the inoculation loop to pick up some of

the bacteria from a starter plate. You will then move the inoculation loop holding the bacteria

into the +plasmid tube. Then gently stir up the inoculation tube to ensure that all of the bacteria

has dispersed. Then use a different inoculation loop in order to not cross-contaminate. Then do

the same thing except now place the inoculation loop into the -plasmid tube. Once you have

obtained the plasmid solution you will need to remove 10uL of the plasmid solution and place it

into the +plasmid microcentrifuge tube using a pipette. Make sure you tap the microcentrifuge to

mix the plasmids into the solution. Then place this solution on ice for 10 more minutes. After

this label each of the 4 agar plates, you should have. Name them, no pGLO LB only, no pGLO

LB/AMP, pGLO LB/AMP, and pGLO LB/AMP/ARA. After the ten minutes are up you want to
 Hinckley 40

take the microfuge tubes out of the ice then gently place them in the water on a floater for 50

seconds. After the time is up take the tubes and place them back in ice for 2 minutes. After that,

remove the tubes from the ice and place them on a workbench where you will then add 250uL of

LB broth to the +plasmid tube and gently tap the tube to mix the solutions. Repeat this step for

the -plasmid tube and incubate both the + and - plastic tubes for 10 minutes at room temperature.

Using a different pipette from before transfer 100uL of the + plasmid microcentrifuge and the -

plasmid microcentrifuge on an appropriate plate. Make sure to use a different pipette for each

plate in order to prevent cross-contamination. Finally, use an inoculation loop in order to spread

the mixture over the plate. Yet again make sure to use a different loop for each plate. Lastly,

incubate the plates at 37 degrees celsius in an incubator for 24 hours, then analyze the results.

Data Collection and Analysis:

No UV Light Under UV Light

 Hinckley 41

Discussion and Conclusion:

Discussion: After close examination of this experiment. I have found that the bacterial plates

that contained the pGLO had the ability to grow even within the presence of AMP, a substance

that is meant to prevent bacteria from growing. For example, the E coli, mixed with the only LB

plate, died after exposure to AMP; this plate also lacked pGLO. However, in both of the plates

that contained pGLO and AMP, there was still bacterial growth and these cultures continued to

thrive. Another thing that was examined closely was the appearance of ARA in the pGLO

LB/AMP/ARA. Additionally, under a UV light, this plate glows in the dark. I have found that

this is due to transformation in which the bacteria take in the plasmid for ARA which will then

express the gene for glowing. Overall, the LB only died, the pGLO LB/AMP survived, and only

the LB/AMP/ARA glowed under UV light. Of course, like any experiment, there can be multiple

different types of results. One error that could have occurred is that the same innocuous loop was

used more than once on the DNA which would lead to cross-contamination and completely

wrong results. Another error that could have occurred is that the pipette was reused which would

yet again cause cross-contamination. Another error could occur due to mislabeling and the LB

with AMP that didn’t contain pGLO could be mislabeled with the LB/AMP containing pGLO

this would make the results confusing. Therefore the results would not be accurate.

Conclusion: In conclusion, these laboratory experiments support my hypothesis. I predicted that

the bacteria that contains the pGLO plasmid will be able to produce bacteria that contains a

resistance to AMP compared to the bacteria without the pGLO plasmid. Not only this but the

LB/AMP/ARA bacteria was also able to produce more bacterial colonies. For example, only the

plates containing the pGLO combined E Coli was able to keep reproducing. Before discussing

improvements that could have been made to this experiment I would like to discuss questions
 Hinckley 42

posed by this experiment. First and foremost the pGLO plasmid is composed of 3 different types

of genes, the araC gene, the GFP gene, and the pAMP gene. Also, the CaCl2 was added to the

solution in order to help open up the plasma membrane on the bacteria so this will allow the

bacteria to take in the plasmid, allowing it to transform. Lastly, E. coli is not naturally resistant to

E coli because, without the pGLO plasmid, the bacteria colony would thrive or survive in AMP.

Now, I have a couple of suggestions for this experiment. First of all, during this experiment, it is

extremely simple to cross-contaminate DNA samples, so I would recommend only using three

plates instead of 4 because it would lower the chance of contamination. Lastly, the heat of an

incubator can drastically increase an increase which can change results so I recommend allowing

the plates to go on a heat plate so there is an even heat distribution.

Questions: There are numerous ways in which this laboratory experiment can be continued. One

way in which it can be continued is to include a plasmid different from pGLO that will pertain to

different results. Another way can be adding multiple different types of bacteria within the LB

bacteria this might show if bacteria are selective when choosing different genes. Then on top of

this experiment, the AMP can be added which could also show if some plasmids can become

resistant when mixed with other plasmids. In addition to this another substance could be used in

order to open the plasmid other than CaCl2

This investigation has raised a few questions. First and foremost, what would happen if

CaCl2 wasn’t used? What would happen if E. coli was exposed to multiple different types of

plasmids at once? Would the E. coli have preferred a certain plasmid over the other? If so, would

this plasmid be the most beneficial to the E coli’s survival?

 Hinckley 43

Paige Hinckley Lab Write Up #6 2/17/2023

NaCl Effect on the growth and Development of Brine Shrimp

Abstract: During this laboratory experiment, I placed four different salinity levels into 4 Petri

dishes then using a fine paintbrush I placed the brine shrimp cysts into each solution which

contained dechlorinated water and NaCl. The first petri dish contained a NaCl level of 0%. The

second dish contained 2% of NaCl. The third contained 6% of NaCl and the fourth contained 8%

of NaCl. These dishes were then transferred to microscope slides with double-sided tape on each

side. The number of hatched and dead brine shrimp were examined at 0 hours, 24 hours, and then

48 hours. Overall, the main goal of this experiment was to see how salinity levels affect hatching

viability.

Introduction: A question was posed during this experiment; how do salinity levels affect the

hatching viability of brine shrimp? Brine shrimp are small organisms often referred to as “sea

monkeys.” These small organisms thrive in salty environments, more specifically the Great Salt

Lake of Utah. Brine shrimp contain a total of 14-17 steps within their lifecycle, however, the

main steps are; Cysts, Nauplii, Juveniles, and adults. The main stage we’ll focus on during this

experiment is the cyst stage. Cysts are just like eggs in the sense that they contain the brine

shrimp embryo within them. Cysts also have a unique way to survive when conditions are not

ideal. The mother brine shrimp will release dormant cysts which are arrested in development

until conditions are ideal. In this experiment, we will be focussing on the salinity of the brine

shrimp environment. Salinity means the salt concentration within the water and can be a large

determining factor in the growth and survival of all organisms. For example, let's say an ocean
 Hinckley 44

contained a NaCl concentration of 4% then a human was to drink that water and their cells

contained a lower NaCl concentration then the cells would be placed in a hypertonic solution

meaning that salinity in the outside environment is higher than the inside leading to the human

cells shriveling up and eventually dying. On the other hand, if a person drank too much pure

water their cells would be in a hypotonic environment meaning that water would flow into the

cells causing the cells to swell and lyse which can also lead to death. Therefore the perfect

tonicity would be an isotonic environment, this is when materials are being exchanged out of the

cell at the same time. With all of this being said, the brine shrimp, in this case, would survive in

an environment that is best suited to their needs and the best will come from an isotonic

environment. A null hypothesis for this experiment would be that Environmental salinity levels

within the environment will not impact the growth and development of brine shrimp. I predict

that the brine shrimp with a salinity of 8% NaCl will have higher hatching viability than the

lower NaCl concentrations.

Materials and Procedure:

Materials: Before starting this experiment you will need some supplies. For starters, you will

need brine shrimp cysts, these can commonly be found online and are usually marketed as “seas

monkeys.” Next, you will need NaCl which is commonly known as table salt, this will also be a

factor for salinity during this experiment. Additionally, 4 beakers are needed in order to hold the

NaCl solution. You will need a scale and weigh boats to measure the amount of NaCl. In order to

place the NaCl solution on the weight boats you need a lab spoon or normal spoon which can be

found at your local grocery store. You will also need a graduated cylinder in order to measure the

amount of water you are using. Next, a stirring rod is needed in order to mix the solution. Next,
 Hinckley 45

you will need pipettes, these can be disposed of when you are finished since they are difficult to

clear. Petri dishes are also needed and will hold the solution once it is finished. Now, you will

need dechlorinated water which acts as the base for the NaCl solution. Additionally, 4

microscope slides will be needed, these will be the holder for the solution in addition to the brine

shrimp eggs. In order to transfer the brine shrimp cysts to a microscope slide a fine paintbrush

will be needed. Double-sided tape will be needed to line the microscope slides and you should

have a pair of scissors handy in order to cut the tape. Lastly, a semi-permanent marker will be

needed in order to label the microscope slides so that the solutions do not get mixed up.

However, don’t forget safety. You should wear goggles, a lab coat, and gloves, and if you have

long hair you need a hair tie. In order to get rid of the material once the experiment is completed

use Hazard Waste Disposable bags.

Procedure: Once you have collected all of the materials it is time to begin the experiment. First

and foremost, gather 4 beakers and label them 0%, 2%, 4%, and 8%. Then in order to make the

saline solution for the 0% all you have to do is put 100 ml of dechlorinated water into a beaker.

Then to make the 2% for the rest of the NaCl solutions mixed the right amount of NaCl in

dechlorinated water to make these solutions. Then place 30 ml of NaCl solution into each petri

dish using a pipette. Don’t forget to alternate pipettes after each use. Then place small strips of

double-sided tape onto the 4 microscope slides. Then use a paintbrush to collect brine shrimp

cysts from a brine shrimp bag. Then place each of the microscope slides under a microscope and

count the number of cysts for each solution. Then after 24 hours check the slides again and count

the number of cysts, hatched brine shrimp, and dead cysts. In addition, remove the dead cysts.

Then do the same thing again after 48 hours. Finally, collect the data within a chart.
 Hinckley 46

Data Collection and Analysis:

% Hatching Viability at Each Salt Concentration

Group 0% 2% 4% 8%

Your DATA 0 25 30 38.1

A 0 22 31 40

B 1 21 34 41

C 0 24 33 42

D 2 22 30 41

E 1 24 31 40

Mean .667 23 31.5 40.35

STDEV .816 2.4 1.64 1.33

2 SEM .667 1.96 1.34 1.09

0 Hrs 24 Hrs 48 Hrs

% # # # Dead or # # Dead or # %
# Cysts
NaCl Cysts Cysts partially swimming partially swimmi Hatching
 Hinckley 47

hatched hatched ng Viability

0 23 18 5 0 13 5 0 0

2 20 16 2 2 11 2 3 25%

4 20 17 1 2 12 1 4 30%

8 21 18 0 3 13 0 5 38.1%

(THE ERROR BARS ARE ON THE VIRTUAL LAB, THEY WOULDN” T

TRANSLATE TO GOOGLE DOCS FROM GOOGLE SLIDES)

Discussion and Conclusion:

Discussion: After close examination of this experiment, I have found that brine shrimp cysts that

were in the solution with a NaCl concentration of 8% had higher hatching viability than the rest
 Hinckley 48

of the NaCl concentrations. During the experiment the 0% NaCl solution caused 10 cysts to die.

This is partially due to the fact that the cysts were submerged within a hypotonic environment.

Meaning that the cysts that are most likely swelled and lysed, eventually causing them to pop

and die. In the NaCl concentration of 2%, some cysts died while others survived which is

partially due to instability within the environment which is the same for the NaCl concentration

of 4%. However, the NaCl solute of 8% was the perfect environment for the brine shrimp cysts

and it was an isotonic solution meaning that the cyst and brine shrimp could thrive. Of course,

like any experiment, there can be multiple different types of errors. One error that could have

occurred is that the brine shrimp cysts could have already been dead or squished in the bag

which could cause them to not hatch leading to results that would be wrong since no brine

shrimp cysts would hatch. Another possible error is that the NaCl solutions were oversaturated

and instead of the salinity being 8% the NaCl concentration would be 12% which could lead to

the cysts shriveling up and dying. Additionally, the beakers could be wrongly labeled leading to

mix-match results that do not add up correctly. In all of these error instances the inaccurate

results could lead to a demolished experiment.

Conclusion: In conclusion, this laboratory experiment supported my hypothesis. I predicted that

the brine shrimp with a salinity of 8% NaCl will have higher hatching viability than the lower

NaCl concentrations. Through close examination, I have found that brine shrimp cyst hatch and

are able to survive in the 8% NaCl solution meaning that a higher salt concentration is better for

the growth and development of brine shrimp. For example, throughout the entirety of the

experiment, the 8% NaCl solution had no dead cysts. However, there were 8 swimming at the

end of the 48 hours and the hatching viability was 38.1%. On the other hand, the 0% NaCl

caused the death of 10 cysts and has a hatching viability of 0%. In addition, the NaCl solution of
 Hinckley 49

4% has a hatching viability of 30% and the NaCl solution of 2% has a hatching viability of 25%.

Therefore the NaCl solution of 8% will allow for the survival and development of brine shrimp.

Now, I have a couple of suggestions for this experiment. First and foremost, this experiment

could use a pre-made salt solution so that the salt concentration couldn’t be mixed up causing

weird results. Secondly, the cysts should have been examined for a longer period of time because

there is a possibility of a few cysts hatching later on and some cysts take longer than others.

Lastly, I recommend that the number of cysts should be carefully measured to be the same

amount so that the rate of the cysts hatching can not be due to the number of cysts.

Questions: There are numerous ways in which this laboratory experiment can be continued. One

way in which it can be concluded is by using orbeez. Orbeez is just like cysts in a way because

they are originally shriveled up and they will then swell when exposed to water. Then it would be

interesting to see what would happen in different salinities. In addition, the brine shrimp cysts

could be placed into a different solution, containing various substances and concentrations. It

would also be interesting to hydrate the cysts and then place them in the water again to see if the

cysts will become dormant again or if they would die.

This investigation has raised a few questions. First of all, what would happen if the cysts

were placed in a solution of 50% salt, would they shrink even more? Would the outer layer of the

cyst be destroyed? What would happen if a different solution was used in place of the

dechlorinated water? What would happen if the cysts were observed for a longer period of time?

Can brine shrimp survive in the same environment as cysts that have entered a dormant state?
 Hinckley 50

Paige Hinckley Lab Write-Up #7 3/2/2023

A Common Ancestry Between Organisms

Abstract: During this laboratory experiment I did two different experiments. First, I tested 6

different species of animals and compared a human ADH6 amino acid sequence within a BLAST

program which searches throughout the entire internet to find genetic matches and the data can

change over time. Out of the 6 different types of species I have tested I have found that frogs do

not contain the ADH6 amino acid, meaning that they are not related to humans. On the other

hand, chimpanzees had the highest relation to humans and were 96.52% identical. During the

next experiment, I tested cytochrome P450 2D14 isoform 1X in horses. I found that horseflies

had no ancestry with horses and then I found that zebras had the highest similarities with horses.

With that being said, the main goal of this experiment was to find evidence of common ancestry

as well as evolutionary evidence stating that humans have evolved from other species of

organisms.

Introduction: A question was posed during this experiment; do organisms share common

ancestors? This question was then answered by both the experiment involving humans and

ADH6 amino acid sequences as well as cytochrome P450 2D14 isoform 1X in horses. First and

foremost, all organisms have a common ancestor. A common ancestor is a recent ancestor that

can share similar characteristics as the other organism. Structural evidence indicates common

ancestry, the structural evidence is called homologous structures which are characteristics that

are similar in two species. However, analogous structures also exist and do not represent a

common ancestor even though an organism may look alike. This is why this experiment must be
 Hinckley 51

done in order to show a common ancestor within humans as well as horses. In addition to this

phylogenetic trees as well as cladograms are able to show common ancestry but with a diagram.

Phylogenetic trees are diagrams that represent the evolutionary history of a group of organisms

using fossils. Cladograms are similar to phylogenetic trees; however, cladograms contain lines

that represent a lineage, and branching points that represent nodes, as well as clades. In

cladograms, the common ancestor is the root of all species. In this scenario, the humans were

being compared to 6 other species in order to test common ancestry and to see what animals the

humans came from first. The species were frogs, cows, rats, chickens, chimpanzees, and rhesus

monkeys. For the experiment using the horse, I used 4 other species which were; horseflies,

alligators, zebras, and squirrels. A null hypothesis for this experiment is that organisms do not

have or share common ancestors, therefore, refuting the idea of evolution. I predict that humans

will be most closely related to chimpanzees and horses will be most related to zebras. This will

then support the idea of common ancestry and evolution.

Materials and Procedures:

Materials: Not many materials are needed during this procedure. For starters, you will need a

laptop that can run programs and functions well. In addition to this, you will need a pen and

paper to record the results of your experiment. This is all you will need to do. Don’t worry about

any safety gear or cleanup since this lab doesn’t require that.

Procedure: Once you have your computer you are ready for procedure 1. For starters, go to

www.ncbi.nlm.nih.gov. There will be a menu that appears on the right-hand side of the page, you

will want to select BLAST. Once you click on blast there will be 4 options; nucleotide BLAST,
 Hinckley 52

Protein BLAST, blastx, and tblastn. You should click on the protein BLAST. Then copy and past

the ADH6 amino acid sequence which is (>AAH39065.1 ADH6 protein [Homo sapiens]

MSTTGQVIRCKAAILWKPGAPFSIEEVEVAPPKAKEVRIKVVATGLCGTEMKVLGSKHLD

LLYPTILGHEGAGIVESIGEGVSTVKPGDKVITLFLPQCGECTSCLNSEGNFCIQFKQSKTQ

LMSDGTSRFTCKGKSIYHFGNTSTFCEYTVIKEISVAKIDAVAPLEKVCLISCGFSTGFGAA

INTAKVTPGSTCAVFGLGGVGLSVVMGCKAAGAARIIGVDVNKEKFKKAQELGATECL

NPQDLKKPIQEVLFDMTDAGIDFCFEAIGNLDVLAGRA DSTSLNWLLIIWQRS). Then

type in the organisms box one of the 6 organisms. Make sure the database is set to Reference

Proteins (RefSeq protein). Then make sure the program selection is blastp. You will then click

the BLAST button, this will take a while to load all of your information. After you are done with

this see if the ADH6 gene is present or absent. If it is present add the query cover, e-value, and %

identical. Repeat this step for all 6 species. Then in procedure 2 go to the same website. You will

then type in your desired protein or gene at the top of the search bar. Then a new section will

show up while showing genes and protein. Depending on what you have, choose what category

your selected protein or gene falls into. Then add your animal's name to the search bar along

with the protein/gene and hit search. Then hit FASTA which will show up and give you the

sequence that you need to BLASt the protein/gene and find a common ancestor. Then do the

name thing that you did in the first procedure using BLAST for 4 other species of your choice.

Lastly, create a cladogram for both procedure 1 and procedure 2.

Data Collection and Analysis:

 Hinckley 53

Procedure 1:

Organism Present/Absent Query Cover E-Value % Identical

Humans --- --- --- ---

Cows Present 93% 6e-162 79.50%

Rats Present 93% 3e-138 67.03%

Chimpanzees Present 97% 0 96.52%

Chickens Present 96% 4e-132 63.54%

Rhesus Monkey Present 77% 2e-79 90.44%

Frogs Absent --- --- ---

Procedure 2:

Organism Present/Absent Query Cover E-Value % Identical

 Hinckley 54

Horses --- --- --- ---

Squirrels Present 100% 0 76.33%

Plains Zebra Present 100% 0 88.36%

Alligators Present 99% 0 55.51%

Horseflies Absent --- --- ---

Discussion and Conclusion:

Discussion: After close examination of this experiment, I have found that both horses and

humans have common ancestors and Horses are most closely related to zebras while humans are

most closely related to chimpanzees. During this experiment, cows were 79.50% identical, rats

were 67.03%, chimpanzees were 96.52%, chickens were 63.54%, rhesus monkeys were 90.44%,
 Hinckley 55

and frogs were not related to humans at all. With this being said, since the chimpanzees were the

most similar to humans it means that humans common ancestry is chimpanzees. However,

humans have no relation to frogs meaning that frogs are not a common ancestor. During

procedure 2 squirrels were 76.33% identical to horses, plain zebras were 88.36%, alligators were

55.51%, and horseplies were not related to horses at all. These results show that the most

common ancestor is a zebra in this case while horses do not share ancestry with horse

flies.shrimp cysts and it was an isotonic solution meaning that the cyst and brine shrimp could

thrive. Of course, like any experiment, there can be multiple different types of errors. One error

that could have occurred is that the ADH6 amino acid sequence could have been typed

incorrectly within the system which would make the results unusable from this experiment.

Another error that could have occurred is that the site was having a glitch which made it record

false answers. Next, I could have recorded a different protein sequence in both the 1st and 2nd

procedure because each protein has different types of subscripts that can make this experiment

confusing. In all of these error instances the inaccurate results could lead to a demolished

experiment.

Conclusion: In conclusion, this laboratory experiment supported my hypothesis. I predicted that

humans will be most closely related to chimpanzees and horses will be most related to zebras.

Which will then support the idea of common ancestry and evolution. Through close examination,

I have found that chimpanzees are a common ancestor of humans and zebras are a common

ancestor of horses. For example, throughout the entirety of this experiment chimpanzees had a

high similarity of 96.52% to humans in addition to this with the cladogram the chimpanzees were

the closest to humans out of all the other 6 species. In addition to this during procedure 2, the
 Hinckley 56

zebras were 88.36% related to horses which was the highest out of all of the other 4 species.

Also, the cladogram shows common ancestry between the zebra and horse. Now, I have a couple

of suggestions for this experiment. First and foremost, this experiment could have an easier

access code to each protein which would decrease the present for error. Secondly, there could not

have been enough organisms tested and if there were you could find that there are other

organisms more closely related to the organisms stated. Lastly, I recommend that other

organisms could be used such as plants and bacteria to show relations and common ancestor

between all organisms.

Questions: There are numerous ways in which this laboratory experiment can be continued. One

way in which it can be concluded is by using other organisms such as fungi, bacteria, and plants

since we have already tested animals. In addition, the plants and other organisms could be

examined by using the same exact BLAST method.

This investigation has raised a few questions. First of all, what would happen if bacteria

were placed in the BLAST system? Next, is it possible for two separate organisms to be 100%

identical without asexual reproduction? Did habitats play a role in the proteins' existence?

Cited Sources:

“Protein BLAST: search protein databases using a protein query.” Protein BLAST: search protein

databases using a protein query,

https://blast.ncbi.nlm.nih.gov/Blast.cgi?PROGRAM=blastp&PAGE_TYPE=BlastSearch

&LINK_LOC=blasthome. Accessed 2 March 2023.

 Hinckley 57

Paige Hinckley Lab Write-Up #8 3/22/2023

Isopod Reactions to Different Environments which includes the

experimentation between a Wet vs. Dry environment as well as an Acidic vs. Basic Environment.

Abstract: During this laboratory experiment I tested two different experiments. First, I placed

ten isopods into a choice chamber for the wet vs. dry experiment. At the 1st minute mark, 7/10 of

the isopods preferred the wet environment and when the ten minutes was up 100% of the isopods

picked the wet environment. Then I placed 10 isopods in an area with one-sided diluted

hydrochloric acid and the other empty. During the one-minute mark 4/10 of the isopods preferred

the acidic environment and when the ten minutes were up 6/10 preferred the acidic environment

meaning that there wasn’t any preference. Overall, the main goal of this experiment was to

determine which environments isopods preferred.

Introduction: A question was posed during this experiment; which environments do isopods

prefer, do they prefer a dry environment over a wet one? Or do they prefer an acidic environment

over a basic one? Isopods go under a plethora of different names but they are most commonly

referred to as rolly pollies. Isopods usually enjoy a habitat that consists of leaf cover, heavy

organic mulch, and rocks. In many cases, isopods prefer soil that is composed of organic matter

such as feces and decomposed animals and stumps because these materials are good for

nutrient-rich soil. However, isopods are usually disastrously affected when the soil is too wet,

tilled, and has an acidic pH. Moreover, isopods are diurnal which means that they prefer to stay

awake during the day and will sleep at night. Also, Isopods are a detriment, meaning that they
 Hinckley 58

eat waste detritus, which is organic matter, for energy. In addition to this, Isopods have a very

special way of protecting themselves. When isopods feel threatened they will curl up into a ball

in order to protect themselves. The proximate cause, how something happens, is that something

is disturbing the isopod which acts as a stimulus. Then the ultimate cause, of why something

occurs, is that this action increases the isopod's chance of survival and can protect them against

predators which increases the overall fitness, the ability to survive and reproduce, of isopods. In

this experiment, we will be focussing on how different environments impact isopods. For

example, the wet vs. dry environment could represent the environment in which isopods like to

live because a wet environment is usually depicted by a decent amount of annual rainfall.

Additionally, the acidic vs basic environment could represent the preferred soil pH which can

vary across the world. You may be wondering, what is pH? Well, pH stands for potential

hydrogen, meaning that if something has a higher H+ concentration it will be more acidic

compared to something with a high OH- concentration which would be considered basic. A null

hypothesis for this experiment would be that there would be no difference in which type of soil

environment the isopod will prefer. I Predict that isopods will prefer a wet environment over a dry

environment and that the isopods will prefer soil of a basic pH over an acidic pH.

Materials and Procedures:

Materials: Before beginning this experiment you will need some supplies. For starters, you will

need 20 isopods also known as roly-polys which can be found at a pet store or found outside

under rocks and deep within the soil. Next, you will need Petri dishes in order to separate the

isopods into 10 isopods in each dish. In order to transfer to the Petri dishes you will need a

fine-tipped paint brush which can be found at your local arts and crafts store. Then, you will

need two choices: dishes that are important for the experiment in order to test the preferences of
 Hinckley 59

the isopods. Now for the basic vs. acidic lab, you will need regular water which will be the basic,

which will also be used during the wet lab, as well as 1% diluted hydrochloric acid for the acidic

side. Now, pipettes will also be used, these pipettes can be normal plastic ones that should be

disposed of at every use to prevent contamination. Additionally, filter paper will be needed

which will be used to hold the liquid solutions for this experiment. Lastly, you should have a

timer on hand which can either be a phone timer or a stopwatch, and make sure it can go up to 10

minutes. However, don’t forget safety. You should wear goggles, a lab coat, and gloves, and if

you have long hair you need a hair tie. In order to get rid of the material once the experiment is

completed use Hazard Waste Disposable bags.

Procedure: Once you have collected all of the materials it is time to begin the experiment. First

and foremost, gather two Petri dishes and separate a total of 20 isopods into each petri dish so

each petri dish should contain about 10 isopods. Then gather two choice chambers and place the

filter paper on each side of the choice chamber. Now, for the Wet vs. Dry experiments place 5-10

drops of water onto the filter paper on one side while leaving the other side dry. For the acidic vs.

basic experiment drop 5-10 drops of dilute hydrochloric acid onto one side of the other choice

chamber all the while keeping one blank. Then gently add the isopods to each choice chamber by

using a paintbrush which will ensure safe and stress-free transfer for the isopods. Now, for both

of these experiments count the number of isopods in each environment over 1-minute intervals,

and then at 10 minutes stop counting and that will be the end of your observed results. After that,

find the mean of each environment and you will have to calculate the Chi-square value between

expected and observed results.

Data Collection and Analysis:

 Hinckley 60

Data Chart One:

Minutes # in Wet Environment # in Dry Environment

1 7 3

2 7 3

3 6 4

4 7 3

5 8 2

6 7 3

7 10 0

8 10 0

9 10 0

10 10 0
 Hinckley 61

Chi-Square:

Wet Environment Dry Environment Total

Observed-O 8.2 1.8 10

Expected- E 5 5 10

O-E 3.2 -3.2

(O-E)2 10.24 10.24

(O-E)2/E 2.05 2.05 4.10

 Hinckley 62

Data Chart Two:

Minutes # in acidic environment # in basic environment

1 4 6

2 3 7

3 3 7

4 4 6

5 4 6

6 6 4

7 7 3

8 8 2

9 7 3

10 6 4
 Hinckley 63

Chi-Square:

Acidic Environment Basic Environment Total

Observed-O 5.2 4.8 10

Expected- E 5 5 10

O-E .2 -.2

(O-E)2 .04 .04

(O-E)2/E .008 .008 .016

 Hinckley 64

Procedure 1:

Discussion and Conclusion:

Discussion: After close examination of this experiment, I have found that isopods prefer a wet

environment over a dry one and do not have a preference when it comes to soil pH. During the

wet vs. dry experiment the isopods have shown to favor a more moist environment because when

the during the one-minute mark the 7 isopods were in the moist environment while 3 isopods

were in the dry environment, then at the ten-minute mark 10 of the isopods were in the moist

environment while 0 were in the dry area. On top of this after examination of the Chi-Square the

calculated amount, 4.10 was greater than the degrees of freedom (3.84) meaning that it is true

that isopods prefer a wet environment since the null hypothesis that the difference between the

expected and observed results is due to chance. In addition to this during the acidic vs. basic lab

at the one-minute mark, there were 4 isopods in the acidic environment and 6 in the basic. Then

at the ten-minute mark, there were 6 isopods in the acidic environment and 4 isopods in the base.

This shows that the isopods truly do not care if their environment has an acidic or basic pH.

Additionally, when the Chi-square was completed the calculated value (.016) was less than the

degrees of freedom value (3.84) which means that the null hypothesis of the difference between

observed and expected results is due to change. Overall, this shows that isopods prefer a wet

environment over a dry one and they do not have a pH preference when it comes to soil. Of

course, like any experiment, there can be multiple different types of errors. One error that could

have occurred is that too much water was added during the wet vs. Dry lab making the isopods

stray away from a puddle of water that could kill them. Another error that could have occurred is

that during the acidic vs. basic lab, the pH could be on completely opposite ends of the spectrum

and some substances could have been a little too acidic. Additionally, the isopods could have
 Hinckley 65

been stressed out from transportation and didn’t want to move at all or might have died but

seemed to be alive.

Conclusion: In conclusion, this laboratory experiment supported and refuted my hypothesis. I

predicted that isopods will prefer a wet environment over a dry environment and that the isopods will

prefer soil of a basic pH over an acidic pH. The idea of isopods preferring a wet environment over a dry

one was correct. For example, when I tested the Chi-square the null hypothesis was correct which can

allude to my alternative hypothesis being correct. However, I was incorrect when I stated that isopods will

have a preference for a pH level that is basic which is completely wrong because this experiment has

shown me that the isopods do not have a preference for pH. For example, when I tested the Chi-square for

this experiment the null hypothesis was accepted meaning that my hypothesis was refuted. Now, I have a

couple of suggestions for this experiment. First and foremost, this experiment should use cotton

instead of filter paper because filter paper creates a puddled mess while cotton swabs hold

moisture. Secondly, more organisms should have been tested during this experiment for

better-proven results as well as more accurate ones. Lastly, I recommend that the organisms were

kept in quarantine and were in good health before this experiment to ensure that the results

weren’t influenced by the stress of this experiment.

Questions:

There are numerous ways in which this laboratory experiment can be continued. One way in

which it can be concluded is by testing the variables of light and dark during this experiment

with a mixture of wet and dry environments to see if a dark or a light environment more heavily

influences isotopes. In addition, different species of isotopes could be used in order to find out if

the species of isotopes determine their preferences for a certain environment.

 Hinckley 66

This investigation has raised a few questions. First of all, what would happen if you put a

few predators of the isopods that favored the wet side and see if the isopods find the moisture to

be necessary? What would happen if the isopods were placed at different temperatures? What if

we were to create mini-ecosystems, which ecosystem would support the isopods?

Cited Sources:

“pillbug - Armadillidium vulgare.” Entomology and Nematology Department,

https://entnemdept.ufl.edu/creatures/MISC/Armadillidium_vulgare.htm. Accessed 22

March 2023.
 Hinckley 67

Paige Hinckley Lab Write-up #9 March 30, 2023

Partner: Mikayla Lawrence

Using the Mark and Recapture Method in Order to Calculate the Size of a Population

Abstract: During this laboratory experiment IN imitated a laboratory experiment using beans

called the mark and recapture method. First I gathered a handful of beans from a bag, counted

them, and then replaced them with darker-colored beans to represent marked individuals. This

process was then repeated three more times. Through these results, I calculated that the

population size was about 1056 individuals while my estimated population size was only 321.

Overall, the main objective of this experiment was to see the effectiveness of the mark and

recapture method and how a scientist is able to estimate the population size without counting

every single individual within a population.

Introduction: A question was posed during this experiment; is the mark and recapture method a

good and effective way to estimate the population size of wildlife? The mark and recapture

technique is utilized by a plethora of scientists all over the world and is used to estimate the

population sizes of certain species without scientists needing to meticulously count each

individual organism of said species. Scientists use this technique by capturing a bunch of

individuals and then placing harmless tags on them, this number is then recorded. Then scientists

will continuously collect the same species and write down the organisms they mark as Mi which

will increase after each capture period. Then the captured amount would be recorded as Ci and

finally, the number of marked individuals collected would be recorded as Ri. With this method,

scientists are finally able to get a good idea of how large a population is. Additionally, the
 Hinckley 68

number of individuals within a population can be calculated by N=MC/R. For example, I

estimated that the population size would be 321 beans while using the mark and recapture

method the population size was 1056 which is a significant difference. Then when I counted the

number of individuals one by one it turns out that there were 7921 individuals. Even Though the

difference is extremely broad the mark and recapture method would be effective it just needs to

go through a series of different rounds. A null hypothesis for this experiment would be that the

mark and recapture method would have no difference in accuracy within the estimated

population size. I Predict that the mark and recapture method will be more accurate than the estimated

population method.

Materials and Procedures:

Materials: Before beginning this experiment you will need a few supplies. First and foremost,

you will need a gallon-sized plastic or paper bag in order to buy your bean which will simulate

an environment, this can be found in your local grocery store. Then a measuring cup is needed in

order to scoop the beans from the large bag. More importantly, two different types of beans will

be needed and these will represent the population. Make sure that the two different beans are

about the same shape and size however, they should be different colors. The light beans should

symbolize the population while the dark-colored beans will represent the captured organisms

within the population that have already been marked. Thankfully, no safety equipment will be

needed for this experiment.

Procedure: Once you have collected all of the materials it is time to begin the experiment. First

and foremost, measure two cups of the light-colored beans and place them in the bag but do not

count them yet, then make an educated guess to see how many beans you have collected. Then

use your hand and grab a handful of light-colored beans and place them out on your desk. The
 Hinckley 69

group captured will represent the first group of captured individuals (C1), so then count these

beans and record the number. Now in order to mark the beans that have been captured simply

replace the light-colored beans with the darker-colored ones. This number of beans will now be

the number of marked individuals which will be the M2 value on your chart. Then return the

marked beans to the bag of light-colored beans and shake it well. Then without looking grab

another handful of beans from the bag which will represent the second capture group of

individuals. Then count the number of beans that you have grabbed which will be the C2 value.

Now, in order for you to examine the beans that you have collected you will need to count the

number of marked individuals that you have captured and write the value as R2 in your table.

Then count that number of unmarked light-colored beans and then mark them by replacing them

with dark-colored beans. Record the number of individuals you just marked under the M2

section of your chart which will be the value for M3. Then, return all the marked beans collected

and place them back into the bag of light-colored beans and shake the bag thoroughly. Yet again

you will need to grab a handful of beans from the bag and record the number of beans that you

collected under C3. Then record the number of marked beans which would be R3 and then count

the number of unmarked individuals and mark them by replacing them with dark-colored beans,

record the number just counted under M3, and record the resulting sum under the M4 value.

Without looking, grab another handful of beans and place them on your desk and record this

number as C4. Then count the number of beans that were marked under R4. Now, use the

equation, M2C2+M3C3…/ R2+R3… in order to calculate for N. Finally, count the actual

amount of beans in your bag and record that in table 1 and separate the beans back into their

respective beakers.
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Data Collection and Analysis:

The mark and Recapture Method using Beans as the Capture Individuals

Sampling # of Marked individuals # of Individuals # of individuals

Time in the Population (Mi) Captured (Ci) Recaptured (Ri)

1 0 132 0

2 132 137 8

3 269 133 43

Estimated Population Size 321

Calculated Population Size 1056

Actual Population Size 7921

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Discussion and Conclusion:

Discussion: After a close examination of this experiment, I have found that the mark and

recapture method is an effective method in order to predict population size in wildlife. During

this experiment when I estimated the population size I believed that it was about 321 beans,

however, after further experimentation, I found that the actual population size was 7,921 which is

a significant difference. Even though the calculated population was still over 6,000 fewer

individuals it was closer to the actual population size than the actual meaning that the mark and

recapture method is still effective. Of course, like any experiment, there can be multiple different

types of errors. One error that could have occurred is that the mark and recapture method could

have been done far too few times which could make the results not as accurate. Another error

that could have occurred is that the counting may have been a problem since there were so many

beans. Lastly, when calculating the N value there could have been errors with numerical inputs
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causing false results. Of Course in all of these error instances the inaccurate results could lead to

a demolished experiment.

Conclusion: In conclusion, this laboratory experiment supported my hypothesis; I predicted that

the mark and recapture method will be more accurate than the estimated population method. For example,

when I performed the capture and release method I got a more accurate result of 1056 for the calculated

population size while when I attempted to estimate the population size I had a completely random guess

of 321 beans. Overall, this experiment supports my hypothesis. Now, I have a couple of suggestions

for this experiment. First and foremost, this experiment should have contained more capture and

release experiments, about 10 more, so that the results are more accurate and I would have

probably gotten a result closer to the actual population size. Secondly, Larger beans could have

been used so that this experiment would take less time so I could focus on more experiments.

Lastly, I recommend that there were more dark-colored beans because I eventually ran out which

prevented me from furthering this laboratory experiment.

Questions:

There are numerous ways in which this laboratory experiment can be continued. One way

in which it can be continued is by using more beans to further the results and create a larger

population. Additionally, this laboratory experiment could be continued in the real world using

organisms from a local stream and using a mark and recapture method to estimate the population

size. Also, A small measuring cup could be used to pick the beans out of the bag because it will

represent the meticulousness of the mark and recapture method.

This investigation has raised a few questions. First of all, what would happen if there

were fewer beans within the population? What would happen if living organisms were used for

this experiment compared to the beans tested? What if we were to mimic the real-world
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conditions of the mark and recapture method and create a laboratory experiment based on that?

Lastly, what would happen if there was a larger variety of beans that represented how many

times the individual had been captured?