Nathan Orlando - Jacob Schmitz

Mr. Estapa
Biology 9A
29 February 2016
Experimental Design
Materials:
1000 mL glass beaker
15 samples of bryopsis
250 mL glass beaker
15-250 mL clear plastic cups
3-2 L bottles
ti nspire
smartphone timer
citric acid
baking soda
scale in g
distilled water
sea salt
tweezers
paper towel
API pH test strips
stirring magnet
Procedure:
1. To determine amount of salt needed, tare weight of 150 mL glass beaker. Add salt
to beaker and examine weight. If above the needed amount, remove salt until level. If
below, add until level. Repeat this until a hundredth of a gram.
2. To stir sample, place stirring magnet in 1000 mL glass beaker, add 1000 mL of
water to beaker, then add salt value for 25 ppt. Stir for 5 minutes.
3. To stir sample, place stirring magnet in 1000 mL glass beaker, add 1000 mL of
water to beaker, then add salt value for 35 ppt. Stir for 5 minutes.
4. To stir sample, place stirring magnet in 1000 mL glass beaker, add 1000 mL of
water to beaker, then add salt value for 35 ppt. Stir for 5 minutes.
5. Following the proper ratio of salt to water (3.8 grams per 150 mL), take one of
three 2 L bottle, fill to 1000 mL with distilled water stirred from beaker with 25 ppt

salinity. Test water with hydrometer to ensure 25 ppt salinity. If water is not exactly 25
ppt salinity, start over until 25 ppt salinity is reached. Label 25 ppt.
6. Following the proper ratio of salt to water (5.8 grams per 150 mL), take two of
three 2 L bottle, fill to top of bottle with water stirred from beaker with 35 ppt salinity.
Test water with hydrometer to ensure 35 ppt salinity. If water is not exactly 35 ppt
salinity, start over until 35 ppt salinity is reached. Label 35 ppt.
7. Following the proper ratio of salt to water (7.5 grams per 150 mL), take three of
three 2 L bottle, fill to 1000 mL with water stirred from beaker with 45 ppt salinity. Test
water with hydrometer to ensure 45 ppt salinity. If water is not exactly 45 ppt salinity,
start over until 45 ppt salinity is reached. Label 45 ppt.
8. For trial 1, take 5 uniform, clear plastic cups that can hold 250 mL water from
materials.
9. Take two of the plastic cups, unused, fill with 25 ppt salinity water from 2 L bottle
to exactly 250 mL water. Label one (-,+), label the other (-,-).
10. Take one of the plastic cups, unused, fill with 35 ppt salinity water from 2 L bottle
to exactly 250 mL water. Label it (S).
11. Take two of the plastic cups, unused, fill with 45 ppt salinity water from 2 L bottle
to exactly 250 mL water. Label one (+,+), label the other (+,-).
12. The second factor of the experiment is the pH balance of the water. So, (-) pH is
7. (S) ph is 8. (+) pH is 9.
13. Take cup (-,-), test pH with testing strips labelled above. if pH reads above 7, then
add citric acid until pH reads 7. If pH reads below 7, then add baking soda until pH reads
7. For each solution, stir for 30 seconds, then check again.
14. Take cup (+,-) test pH with testing strips labelled above. if pH reads above 7, then
add citric acid until pH reads 7. If pH reads below 7, then add baking soda until pH reads
7. For each solution, stir for 30 seconds, then check again.
15. Take cup (S), test pH with testing strips labelled above. if pH reads above 8, then
add citric acid until pH reads 8. If pH reads below 8, then add baking soda until pH reads
8. For each solution, stir for 30 seconds, then check again.
16. Take cup (+,+) test pH with testing strips labelled above. if pH reads above 9, then
add citric acid until pH reads 7. If pH reads below 9, then add baking soda until pH reads
9. For each solution, stir for 30 seconds, then check again.
17. Take cup (-,+) test pH with testing strips labelled above. if pH reads above 9, then
add citric acid until pH reads 7. If pH reads below 9, then add baking soda until pH reads
9. For each solution, stir for 30 seconds, then check again.

18. Label 5 spots in the area you are growing A-E. These should be on a table. Make
sure they get the same sunlight per day, in a greenhouse environment. Keep temperatures
and humidity consistent to ensure accurate results.
19. Label cup (-,+) 1, cup (-,-) 2, cup (S) 3, cup (+,+) 4, and cup (+,-) 5. These
numbers are used for randomization.
20. Use ti nspire to generate a random number between 1-5 using the randint(1,5)
function. Generate until all 5 numbers are chosen. In the correct order, the first number
chosen is tested on spot 1, second on spot 2, third on spot 3, fourth on spot 4, and fifth on
spot 5.
21. Measure the weight of each sample of bryopsis, write down on paper to hundredth
of a gram. Label each sample weight as start weight to their designated cup, (1, 2, 3, 4,
5), and carefully place in center of cup. The current weight will be subtracted from final
weight to determine growth in that environment.
22. Leave samples in greenhouse environment overnight.
23. Day two, inspect samples for any unwanted influences, such as objects in the
water, etc. With the proper environment, these shouldnt be an issue. These will be
factored as noise, which will decrease the accuracy of the experiment.
24. Day three, inspect samples again for any unwanted influences.
25. Test pH of cup 1 with testing strips labelled above.
26. If pH of cup 1 is above 9, use citric acid to lower until pH reads 9. For each
solution, stir for 30 seconds, then check again.
27. If pH of cup 1 is below 9, use baking soda to raise until pH reads 9. For each
solution, stir for 30 seconds, then check again.
28. Test pH of cup 2 with testing strips labelled above.
29. If pH of cup 2 is above 7, use citric acid to lower until pH reads 7. For each
solution, stir for 30 seconds, then check again.
30. If pH of cup 2 is below 7, use baking soda to raise until pH reads 7. For each
solution, stir for 30 seconds, then check again.
31. Test pH of cup 3 with testing strips labelled above.
32. If pH of cup 3 is above 8, use citric acid to lower until pH reads 8. For each
solution, stir for 30 seconds, then check again.
33. If pH of cup 2 is below 8, use baking soda to raise until pH reads 8. For each
solution, stir for 30 seconds, then check again.

34. Test pH of cup 4 with testing strips labelled above.

35. If pH of cup 4 is above 9, use citric acid to lower until pH reads 9. For each
solution, stir for 30 seconds, then check again.
36. If pH of cup 4 is below 9, use baking soda to raise until pH reads 9. For each
solution, stir for 30 seconds, then check again.
37. Test pH of cup 5 with testing strips labelled above.
38. If pH of cup 5 is above 7, use citric acid to lower until pH reads 7. For each
solution, stir for 30 seconds, then check again.
39. If pH of cup 5 is below 7, use baking soda to raise until pH reads 7. For each
solution, stir for 30 seconds, then check again.
40. Day four, inspect samples for any unwanted influences.
41. Repeat steps 24-40 on day 5, Friday.
42. Remove each bryopsis sample carefully with tweezers, place on one strip of paper
towel next to each cup.
43. Tare weight of a cylindrical beaker on scale.
44. Measure the weight of each sample of bryopsis by putting in beaker and on scale,
write down as changed weight to their designated cup, (1, 2, 3, 4, 5).
45. Subtract start weight from changed weight to get each difference in mass.
These numbers will be used in the math portion of this experiment.
46. Repeat steps 1-45 three times, to come up with three sets of five changes in mass
of bryopsis. These are the pieces of data used to determine the effect of each variable.

Diagram:

Figure 1. Experimental Setup

Above is the experiment setup complete with the cups, random spots, and samples of
algae, and paper towel. It is located in the greenhouse.