Biology 220 – The Living Cell Lab Name: ____________________ Writing Guide – Module C Due Date:__________________ In all cases, you should refer to the guidelines provided in Appendix I of your laboratory manual. The instructions below provide additional details about this specific assignment. In this paper, you will follow the style of clinical research articles to describe the context, methods, results and impact of your study. There are two parts to this assignment. Part 1: Draft of methods – due in class during the week of C-2. The draft of your methods should include two pieces. A long-form description of each of the following methods and a true methods section (no more than 1-1.5 double-spaced pages). The goal of writing in both ways is to help you make decisions about what the reader needs and expects in a methods section. Long-form: For each method used, write a paragraph that describes: 1) how the method works and 2) what you used it for. An example from the last module is given below. (5 points) True methods: A true methods section is a condensed description that assumes the reader knows how the technique works, but will not know the details of how you did it or why. You can expect that your reader can make solutions, so there is no need to include volumes. However, the temperatures and times of a reaction will impact the result, so it’s important to give those details. In your draft, try to be thoughtful about which details to omit, which to include and which can be cited from previous work. (5 points, evaluated using the rubric on page 1-11 of your laboratory manual). Examples: Long-form: Light spectrometry is a method used to measure the absorbance of a substance. Light of a specified wavelength is emitted from the source. The light passes through the sample before reaching a detector. The spectrometer calculates the difference between the amount of light emitted and the amount detected to determine how much light was absorbed by the sample. The Beer-Lambert law allows you to convert the absorbance into a concentration. Here, we used spectrometry to determine the concentration of p-nitrophenol produced by an enzyme reaction in order to determine the rate of the reaction. True methods: Several dilutions of p-nitrophenolphosphate (range 1.5mM – 15mM) were created by mixing the substrate with 1M Tris-HCl, pH 8.0. Each dilution was mixed with 1 unit of alkaline phosphatase and the absorbance of the product, p-nitrophenol, was measured at 410nm at 15 second intervals for up to 2 minutes. The absorbance readings were converted to concentrations using the Beer-Lambert Law and an extinction coefficient of 18.5/mM*cm. The concentrations were used to calculate the velocity of each reaction and plotted in Michaelis-Menten curves. Part 2: Due in class during the day of D-1. You will write a 6-8 page (double-spaced paper) describing the context, methods, results and implications of your work. General guidelines for each section can be found in the laboratory manual appendix I. Your paper should include: - - - - An abstract that summarizes each section of the paper An introduction that provides clinical and biological context for the study. You will likely want to discuss the prevalence and clinical aspects of cystic fibrosis, the molecular nature of the disease and it’s relationship to bacterial infections, including P. aeruginosa specifically. You will need to read and cite the previous literature in this field. Use the citations in the laboratory manual to get you started. Do NOT cite the laboratory manual – follow the citations given there. Your introduction should also include a brief description of what you did, what you found and what it means. (~2 pages) A methods section based on the instructions given in part 1. (1-1.5 pages) o DNA isolation o DNA amplification o Enzyme digest and gel electrophoresis o Data analysis and statistics A results section that follows the guidelines given in Module B. (1.5-2 pages) o Labeled Gel o Table comparing genotypes and phenotypes and the associated X2 test. A discussion that connects your results to the information given in the introduction and discusses any limitations of the study or future directions (1 page). List of Literature Cited This assignment is worth 30 points and will be evaluated using the rubric on pages 1-11 and 1-12 of your laboratory manual.
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