Lab 2 Separation and Analysis of Casein and Albumin

Lab Writeup Guidelines and Rubric
Introduction
Many settings including professional and medical laboratories require keeping records in
an official lab notebook. Many students will find themselves in this situation and
therefore, the Biochemical Laboratory Exploration course at Saint Paul College requires
the use of a laboratory notebook for all labs. This document details how to properly
format and make data entries in your notebook throughout the semester.
Formatting
Type up your report with sensible margins and fonts/font sizes. Make sure that each
section (Introduction/Methods/etc) are labeled with a heading so they can be found
easily
Lab writeups
Each lab should include the following:
1. Title of the lab.
2. Date(s) the lab took place
3. Your name and partner(s) name(s) if applicable.
4. Introduction.
a. Give the overall purpose of the experiment
b. include any important definitions and relevant background information
5. Methods.
a. Give enough information so another person could exactly repeat the work
described.
b. In the case where you are following a standard procedure, you can just
reference the procedure.
i. but be sure to make note of any changes to the standard procedure
6. Results.
a. It is very important to record any raw data and observations
b. Show all work for any calculations
i. if you are having difficulty showing math functions with your chosen
word processor, you can always write them on a piece of paper, take
a pic and add the pic to your lab report
c. Any printed graphs/data should be inserted directly into the results section
i. be sure any graphs and tables are properly labeled with figure
numbers, captions, units, axes on graphs are labeled, etc.
d. pictures should have captions explaining what they are depicting
i. any pictures of gels or other results that require interpretation should
have descriptive labels and arrows pointing out relevant/important
features
7. Conclusion.
a. Give a summary paragraph about your results and findings.
b. discuss the implications of your results and spend some time interpreting
any data that you think may be incorrect
c. If you are unsatisfied with your lab results, be sure to include what went
wrong and what steps should be done to get more satisfactory results next
time
In order to get full credit for your lab notebook, please read this document carefully.
Each Lab must be represented in your lab notebook. Each writeup will be worth 10
points. There are 7 writeups so 70 points total.
Example Grading Rubric
Title/Date/Name 0.5 0.25
Introduction 2 1.5 1 0.5
Methods 1.5 1 0.5
Results 2 1.5 1 0.5
Conclusion 2 1.5 1 0.5
Overall Neatness/Quality 2 1.5 1 0.5
Below is an example lab writeup
Photosynthesis Lab: Aerobic Cellular Respiration and Pigmentation
Yara King
Lab Dates: Sept 1st 2020 & Sept. 7th 2020
Labmates: Abed Ellan, Mika Eddovich, Kao Yang
Instructor: Jim Gielissen
Introduction
There are two goals for today’s lab.
First, we want to measure rates of oxygen and carbon dioxide creation/usage by spinach plants in an
enclosed chamber. This will serve to highlight the competing actions of aerobic cellular respiration and
photosynthesis in the leaf cells (primarily the palisade mesophyll cells). We learned that aerobic
cellular respiration is a series of metabolic reactions in which cells can create useful chemical energy
while using up gaseous oxygen and creating carbon dioxide as a waste product. We also learned that
photosynthesis is a series of metabolic reactions in plant chloroplasts that takes up CO2
to create
sugar and gives off O2 as a waste product.
Second, we will run two experiments that help to characterize the presence of primary and accessory
pigments in spinach leaves. The first experiment will serve as a qualitative characterization through
the use of simple chromatography. The second will generation an absorption spectrograph that
reflects total relative absorbance from 400nm to 700nm
Methods
We followed the standard procedure in the provided lab handout. There were three instances where
we made slight alterations. The alterations and the reasons for altering are noted below.
● Procedure 1 step 4
○ Our chamber remained dark for 12 minutes instead of the 10 indicated by the
instructions
■ This step took more time than indicated because we were preparing solutions
for procedure 2. We think that this may have impacted our results. See
conclusion section for more information
● Procedure 3 step 6
○ We produced 12 ml of pigment lysate instead of 10
■ We believe this change was due to a mistaken volume measurement by a group
member, and may have produced a slightly diluted sample. See conclusion for
more information
● Procedure 3 step 15
○ When using the spectrophotometer, our final measurement was greater than 10 times
all previous measurements
■ We believe that this is because the number was improperly dedicated by a
group member. This odd value was noted during the lab and remeasured with
similar numbers that were 1/10th the value. See note in results as well as an
explanation in the conclusion for more information
Results
Figure 1: This graph from procedure 1 shows the change in CO2 concentration in our respiration chamber over
10 minutes. Note:This data was collected by computer, raw data is not listed here.
<—– Yellow pigment, at the same level as highest point of solvent (85 mm to bottom)
<——-Yellow/green pigment (66 mm from loading line to bottom border of pigment)
<——-Bright green pigment (55 mm from loading line to bottom border of pigment)
<——-Olive green pigment (42 mm from loading line to bottom border of pigment)
<———Loading Line
Figure 2: The results of procedure 2 showing our spinach pigment chromatography which indicates four clearly
defined pigment colors
Rf
bottom values of each pigment in figure 2
● Olive green: 42/85 = 0.49
● Bright green: 55/85 = 0.65
● yellow/green: 66/85 = 0.78
● Yellow: 85/85 = 1.0
Figure 3: Procedure 3 graphed data showing intensity from 400nm to 700nm. Raw data was collected by biorad
program and not listed here. Results show a peak at about 435 nm and another at about 670 nm.
Figure 4: For comparison purposes, this is a textbook graph showing peaks at approx 435nm and approx
660nm.
Conclusion
Procedure 1: the results from our procedure generally align with the aerobic cellular respiration and
photosynthesis information we learned about in class. Co2 levels decrease when the leaves are
engaging in photosynthesis and increase when they are engaging in aerobic cellular respiration.
Decrease in the rate of Co2 is associated with the light independent half of photosynthesis. CO2 is
taken up by plants through the stomata and transported to chloroplasts via diffusion In chloroplast
stroma, the CO2 is added to the Calvin-Benson cycle. Our decrease in light environments supports
this. Additionally, the increase of Co2 during the dark phase of the experiment supports what we
learned about regarding the creation of CO2 during glycolysis and the Krebs cycle. One potentially
problematic issue with our experiment is that we were supposed to allow our plants to rest in the
darkened chamber for 10 minutes, but we instead let them rest for 12. It is possible that the slope for
Co2 creation is slightly higher than it should be for this reason because the CO2 levels had an extra 2
min. of accumulation. With our current results, the slope is less-steep than the light chamber slope,
however each slope doesn’t necessarily imply an effect on one another. In order to see if our mistake
had any effect, we would need to repeat the experiment with the corrected time rest period.
Procedure 2: After the instructor informed us that the 4 colors we were able to identify correspond to
the following molecules: beta carotene, xanthophyll, chlorophyll a and chlorophyll b, we hypothesized
that the migration speed of the pigment had something to do with polarity, and that the more polar
pigments would move slower due to hydrogen bonding with the filter paper. Figure 5 below illustrates
why each pigment has different polarities. The polar functional groups, especially the ester and
ketone groups of the chlorophyll pigments create many hydrogen bonding opportunities with the
stationary phase filter paper. The results of this procedure turned out about as we hypothesized. The
polar filter paper interacted with the polar pigments more than the nonpolar pigments. The results in
the nonpolar pigments moving up the filter paper faster, separating them by polarity. As a result, we
found the nonpolar pigment beta-carotene on top, followed by the carotenoid xanthophyll, followed by
chlorophyll a and b respectively.
Figure 5: chemical makeup of each of the pigments listed by the instructor. The polar functional groups are
noted with green circles, while nonpolar are noted with red.
Procedure 3: In our last procedure, we sought to recreate an absorption spectrograph that was
depicted in our textbook (see figures 3 and 4). The graph we constructed had two notable peaks at
435 and 670 nm. We also had a peak at 725 nm, however we believe that it was due to an improperly
dictated number. After we noted the third peak, we suspected that it did not reflect actual data, so we
re-ran the experiment at the nanometer range where we saw the notable data. The peak was not
present. Afer some discussion, we believe that the last data point was misunderstood by a factor of 10
due to a misplaced decimal point.

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