Lab 5

Agenda

• Lab Report Guidelines • Lab 3 Theory and Information • Lab 3 Data Analysis: Plotting

Lab Report Guidelines and Rules

Template of expectations for Lab Report rules provided by previous instructor: Samantha Iverson

General Rules

► Be CLEAR and CONCISE!! ► Length doesn’t always indicate a better report

► Write in the 3rd person ► DO NOT write in the 1st person: no “I”, “ME”, “MY”, “WE”

► With the exception of portions of the introduction and discussion, all writing must be in PAST TENSE with PASSIVE VOICE!!

► Ex. Hydrochloric acid was added in 50uL portions to Unknown AA-01-00 until a pH of 1.5 was reached for a total volume of 3.5mL of HCl added.

► Do not repeat yourself, you don’t need to explain the same thing 5 different ways. ► Pages should be numbered starting AFTER the title page

Report Sections

► Title page

► Abstract

► Introduction

► Materials and Methods

► Results

► Discussion

► Conclusion

► References

► Appendix

► Observations

► Note: not all reports/assignments require every section, please see lab manual to see what sections are required for each report/assignment.

► If you provide sections that are not required, they will not be looked at

► Any missing section will result in full point deductions for that section

► Observations must be stapled to the back of every experiment handed in.

Section: Abstract ► The sole purpose of the abstract is to provide the reader with a summary of the major

accomplishments of the experiment ► Although it appears first in the report, it should be written last because it must indicate a

knowledge of intro, Results, and Discussion ► Do not start with “The purpose of this experiment is….” ► This should be no longer than 200 words (4-5 sentences) ► No equations or references here ► Should include

► Purpose of the experiment ► Brief description of how this was achieved ► Brief explanation of the results (include numerical values) ► What was concluded

Section: Introduction ► Here you provide the reader will ALL background knowledge needed to understand the

experiment ► State the goal of the experiment

► Introduce important methods and why they are used ► Provide knowledge of all relevant theory and concepts ► Start very broad and as you progress, be more specific to the actual experiment

► For example: Amino Acid (AA) titration ► Background on AA

► what is titration

► Indicate what happens to AA as they are titrated (molecularly)

► This section should be filled with in text citations ► The last paragraph should include a hypothesis of what will be seen

Section: Materials and Methods

► This section is used to indicate how the experiment was accomplished and what exact materials were used to achieve the goal.

► This section should not be written as a shopping list or procedure manual but should be written in paragraph form with high organization and in your own words. ► NO commonly used equipment or materials (i.e., tips, pipettors,

glassware, ect.)

► NO common actions (ie. Glassware was washed, tubes were labled, ect.)

► Important information to include (but not limited to) ► Chemical names ► Concentrations (with proper units) ► pH ► Company item was purchased from

► If the chemical used was a mix, you must indicate what is in the mixture as well as each individual concentration

Materials

► 1X PBS Buffer

► Tris/Glycine/SDS Running Buffer

► Laemmli Sample Buffer

► β-mercaptoethanol

► Ready Gel Pre-Cast Gel

► SDS-PAGE Broad Range Molecular Weight Protein Marker

► Coomassie Blue Staining Solution

► Destaining Solution

Methods

1. Prepare Ready Gel Pre-Cast Gels.

2. Mix 20 μL of sample and 20 μL of Sample Buffer.

3. Heat for 100 °C for 5 minutes.

4. Load 20 μL of the samples into the wells.

5. Run the gels at 200 V for 40 minutes.

6. Stain the gels.

7. Destain the gels.

8. Dry the gels.

Ex. Hydrochloric acid (HCl) was added in 50uL portions to Unknown AA-01-00 until a pH of 1.5 was reached for a total volume of 3.5mL of HCl added.

Section: Results

► This section is used to display all results obtained in the experiment

► Start this section with a short paragraph (2-3 sentences) introducing the results and giving a short summary of Materials and Methods ► This is especially important when the report doesn’t require a

full Materials an Methods section ► Results should be portrayed in 3 forms

► Figures ► Tables ► Paragraphs

Section: Results Figures

► Figures must have a specific title, properly labeled axis, a specific legend (if graphing more than one line), and a caption. Any important points on the curve should be labeled (ie pKa, pI, lambda max etc.)

► The caption appears BENEATH the figure and contains the figure number: title, and brief description of important data

► The caption should be a smaller font than the paragraphs showing a clear difference between a caption and text

► When referring to figures in text, use the whole word “figure #” at the beginning of a sentence and the short hand “fig. #” in the middle of a sentence.

► Do not make sentences like: In Figure X, we see that….. Instead, make the point and put (Figure X) at the end of the sentence

Section: Results Good Figure Example

Figure #. Title(same as graph). The caption needs to indicate what is being presented in the figure.

• Figures with captions are stand-alone entities, therefore the reader must be able to understand the figure (and caption) without having to read the rest of the text.

• The title should be specific • The axis are clearly labeled, don’t forget

the units. • Caption is beneath the figure and

contains all important information about the figure (results obtained and how, etc.).The information on the caption MUST BE REPEATED in the results main text.

• Important information is highlighted (equation of the regression line and linearity)

Y=mx+b R2= 0.998

Of what?

Section: Results Tables

► Tables or charts in the results section should indicate important data ► Examples: averages of multiple abs readings, % errors, standard deviations ► Any raw data should appear in tables in the appendix

► Tables should have a number and title ABOVE the table. ► No caption, but you can use a note to indicate any relevant to

understanding the table ► Important data such as wavelength used or pH should be clearly

indicated within the table or title. ► The title should be specific to what data you are showing

Section: Results Proper Table Example

► Table #: Absorbance of varying Concentrations of PNP at 400 nm in pH 10 glycine buffer

Concentration of PNP [mM]

Average Absorbance at 400nM

0 0

0.1 0.1

0.2 0.15

0.3 0.2

0.4 0.25

0.5 0.3

*Note: Raw data of triplicate absorbance measurements can be seen in Appendix C Table 1.

• Table has a specific title • Table number and title appear ABOVE

the table • Units are clearly indicated • Only important data is shown in results • Raw data is shown in the appendix

Section: Results Paragraphs

► Open the results section with a brief description of what was done. ► Result paragraphs should summarize trends seen in both figures and

tables ► Reference the raw date and calculations on the appendix. ► Your paragraphs should also indicate unknown sample number as well

as unknown identity or concentration ► Unknown AA-01-00 is consistent with Histidine ► Unknown Spec-01-00 was found to have a concentration of 10 ug/mL of PNP

► Do not explain why you obtained certain results, just state what the results are ► The why belongs in the discussion

Section: Discussion

► Probably the most important section in the whole report ► Here you are linking the results to the theory and concepts given

in the introductions section ► You are highlighting the full significance of the work here ► Your discussion should:

► Analyze the data obtained ► Mention and explain all results ► Explain any strange data obtained ► Link results obtained to introduction theory and hypothesis

DO NOT JUST RESTATE RESULTS!!!! EXPLAIN THEM!!

You can use outside figures here to aid in any explanations. Just make sure they are treated like any figure (add a caption) and properly cited.

Section: References

► References appear after the discussion but before the appendix, please be aware of the order of the report

► References should be in APA or MLA format (Required by John Jay)

► Please put in the References section which format you are using

► In text citations can be in either APA format (Author, year) or MLA format ([3])

► Period appear AFTER THE IN-TEXT CITATION ► You must have 2 OUTSIDE sources of information

► Outside sources do no include: Biochem textbook, lab manuals, recitation powerpoints, or lecture powerpoints (these can be used as sources, they just don’t count towards the 3 required)

► 2 outside sources should be primary journal articles (not reviews) ► WIKIPEDIA is NOT an acceptable source of information

Section: Appendix

► An appendix appears in EVERY REPORT or ASSIGMENT (even though it isn’t indicated in the syllabus)

► Your appendix contains ► Raw data ► Sample Calculations (fully worked out, not just the equation)

► There must be one sample calculation for every calculation used in the report, including average and standard deviation

► This section should be clearly organized like the rest of the report. Do not just throw information here and expect the instructor to understand what you are talking about

► All figures and tables here have titles and captions as they do in the results. (Just continue with the numbering)

Section: Observations

► Observations are anything that you observe during class ► These observations should be clearly organized and labeled in your

notebook. ► Observations should be attached to the back of the report when

handed in ► Either a photocopy of your notebook pages or the carbon copies are

acceptable