Northern Michigan University

CH 450: BIOCHEMISTRY I
LABORATORY
Fall 2016
Instructors: Suzanne Williams (W, R PM, F): NSF 3121, 227-1068, [email protected]
Lesley Putman (T): NSF 3306, 227-1071, [email protected]
Mark Paulsen (R AM) : NSF 3301A, 227-1064, [email protected]
Text:

Instead of having a lab text, there will be handouts posted on EduCat and web links where you can learn
about various techniques.

Expectations: In this lab we will be getting away from step by step instructions where everything is already
spelled out. The instructions may be more general than you are used to, so you will need to come up with the
specific steps for many of the experiments. It will thus be essential for you to prepare for lab beforehand. The
labs will be done in pairs (except for the first one) but you will switch partners for each different experiment. If
you skip one part of a two part lab, leaving your partner with all the work, you will lose half the points for that
lab.
Schedule: Monday 6-8:50 PM, Tuesday 9-11:50 AM, Wednesday 6-8:50 PM, Thursday 9-11:50 AM, or
Thursday 6-8:50 PM. All labs meet in NSF 2214
Week of 8/22
Week of 8/29
Week of 9/5
Week of 9/12
Week of 9/19
Week of 9/26
Week of 10/3
Week of 10/10
Week of 10/17
Week of 10/24
Week of 10/31
Week of 11/7
Week of 11/14
Week of 11/21
Week of 11/28

Check- in, Safety, Pipetting and Standard Curves

Determination of the pKa for p-Nitrophenol
Properties of Amino Acids
A Comparison of Protein Assays Part I: Biuret Assay
A Comparison of Protein Assays Part II: Lowry and Bradford Assays
Alkaline Phosphatase: Determination of Vmax and KM
Alkaline Phosphatase: Determination of Inhibitor Type
Extraction of Antioxidants from Plant Material

Purification of Acid Phosphatase Part I: Ammonium Sulfate Precipitation

Purification of Acid Phosphatase Part II: Gel Filtration Chromatography
Purification of Acid Phosphatase Part III: SDS-PAGE
Review for lab exam and check-out
Thanksgiving-no labs
Lab Exam

For lab you will need:

Safety goggles
Lock for your drawer
Permanently bound lab notebook (i.e. composition book)
Pen

Lab Notebook:
Each student will have their own notebook. It is unacceptable to have two notebooks: one to use during lab
and one to turn in. It is also unacceptable to record data on a piece of paper and then transfer it into your lab
notebook. Notebook points may be deducted for either of these unacceptable practices. Keeping a lab notebook
is a skill to be developed and one that you will need if you ever do research or work in a lab.
When you come to lab each week, you should have the following in your lab notebook:
All data, results and discussion from last weeks lab.
Parts I, II, and IIIa for current weeks lab. (see below)
Lab notebook grading:
At the beginning of each lab you will receive a check sheet for a few things that should be in your
notebook. You will check off the ones you have and return the check sheet to your instructor. The
check sheets will contribute to your notebook grade.
Your notebook will be collected twice during the semester and graded (scaled to 10 points)
Your notebook will be turned in at end of semester for grading. (20 points)
For the lab exam at the end of the semester you will be able to use your lab notebook, so it is to your advantage to
keep a detailed and organized notebook.
General Lab Notebook Format
Include a Table of Contents at the beginning of the notebook.
Number and date each page.
Use permanent ink.
Never use white-out in a lab notebook, just draw a line through mistakes.
Lab Notebook Format for Each Experiment:
I. Descriptive title and date
II. Introduction
a. Objective or purpose
b. Theory and/or background information
c. References
III. Experimental
a. List materials and reagents. Identify concentrations of solutions and what each is used for.
b. List equipment used: model, make and number
c. Procedure:
This is a record of what you actually did in the lab, so it is best to write it while you are doing
your experiments. However, if you want to outline the procedure ahead of time, leave space so
that you can record any changes you might make, and check off each procedure after you
complete it. Include enough details so that you can refer back to your notebook after a few weeks
and know exactly what you did. The more details you include in your notebook, the easier it will
be to interpret your results and write your report. It also helps avoid mistakes when you take the
time to write down what you did. Record any observations as you go along, such as color change,
cloudiness, etc. Record any calculations you use during the procedure (i.e. making up solutions,
diluting samples, etc..)

IV. Data and Calculations

a. Record all raw data in your notebook. Use tables to record your data whenever possible.
i. Don't record raw data on a piece of paper to be transferred into your notebook later.
ii. Dont record raw data on your computer.
b. Show calculations so you know how you came up with the numbers when you refer back to your
notebook later.
c. Any print-out from an instrument should be labeled and attached to a page in your notebook.
d. When you use a spreadsheet to process your data, print it out and paste it in your notebook. Be sure
to show what calculations were performed to obtain the numbers in each column.
V. Results and Discussion.
a. Attach any graphs that you made to interpret your data. The graph should have a reference to the
page where the data is recorded or where the spreadsheet is attached
b. Summarize your results and discuss what your data mean.
c. Indicate what you concluded from your experiment.
d. Discuss how the experiment went, any problems that occurred or things that you would do
differently next time.
Lab Report:
Lab reports will be due at the beginning of the lab period one week after you complete the experimentation.
Points will be deducted for late labs: 5% if turned in later that same day, then 10% per day after that. Write up
your report as outlined below. Please type, but calculations may be hand-written. The format of the lab report is
designed to be similar to a manuscript that could be submitted for publication. One major difference is having a
statement of objective(s) rather than an introduction. This is just to make it simpler for you, since you already
included an introduction in your notebook.
Abstract: A brief description of what you did, and a summary of your results and conclusions. This section should
be no more than 200 words.
Objective: A statement of what you were trying to accomplish in this lab experiment, i.e. your purpose.
Summary of Procedure: This should be a summary and not a regurgitation of the handout or your lab notebook.
Use a narrative format written in past tense. Do not use tables in this section.
Random examples:
A 1 mM solution of glycine (100 mL) was prepared at pH 9.0.
Pea pods (10 g) were extracted with 100 mL of 1 mM acetate buffer (pH 5.5) using a blender. The crude extract
was filtered through cheese cloth and the filtrate was centrifuged for 15 minutes at 10,000 g.
Note that concentrations of reagents are given but the procedural details are left out.
The Lowry Protein assay was performed as described by Lowry et al. 1. The standard curve was prepared using
bovine serum albumin. It covered the range of 0.02 to 0.4 mg/mL.
1

In your reference section include the complete reference for the Lowry assay.

Results and Discussion: Do not start this section with tables or graphs. This is a narrative section which explains
the results and refers the reader to tables or figures*. Your discussion should include: the meaning of your
results and their implications, how confident you are in the data obtained (were the methods accurate and
precise?), how well the procedures worked, anything that went wrong or seemed questionable. The lab

handouts will give you specifics about what to include in the results and discussion section.
*Tables and Figures: These are the presentations of your results. Do not include "raw" data. For
example, if a table of numbers is graphed to show a straight line, include the graph but not the table of
numbers. The latter is "raw" data and is in your notebook. Be sure to include a descriptive figure legend
below each figure. A graph is considered a figure and should therefore have a figure legend but no title.
Tables have a title that includes the information needed to interpret it. Sometimes tables have footnotes,
but not extensive legends like figures. Graphs should be made on the computer; be sure to label the axes
and include units. This website gives you all the details about how to prepare tables, figures and legends:
http://abacus.bates.edu/~ganderso/biology/resources/writing/HTWtablefigs.html
References: In some lab reports you will want to refer to a published work to support your conclusions or to give
the details of a procedure. In this case, please use a standard format: for example, see
http://library.nmu.edu/guides/userguides/style_apa.htm. If you reference a book, list the specific pages
you used. The lab handout is not to be referenced because it is not published. Neither is your lab
notebook. Only cited references need to be listed.
Questions: Some labs will include questions to answer. Include these at the end of your report. They are worth
points toward your report.
Plagiarism. If you cut and paste text from someone elses report, whether it be your partner or someone in
another lab at another time, this is plagiarism and you will get zero points for the lab report.
Lab Grade:

Point
s
Pipetting and Standard Curves
pKa
Amino acids
Pipetting check
Protein assays
Kinetics
Antioxidants
Acid phosphatase: intro
report
Notebook grading
Final notebook
Lab exam
Discretionary points*
Total points

15
20
20
5
30
30
20
10
40
10
20
25
5
250

* Given by lab instructor for following safety guidelines.

NOTE: If you drop this course, withdraw from school, or quit coming, it is your responsibility to check out
of your lab drawer, even if you never used it, otherwise you will be assessed a fee.
Disability-related services: If you have a need for disability-related accommodations or
services, please inform the Coordinator of Disability Services in the Dean of Students Office at 2101
C. B. Hedgcock Building (227-1700 or [email protected]). Reasonable and effective
accommodations and services will be provided to students if requests are made in a timely manner,
with appropriate documentation, in accordance with federal, state, and University guidelines. (f16)