DNA Melting Report Requirements

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  • Follow the lab report general guidelines.
  • Please do not include your Part 1 report in this final report.
  • Provide a thorough and accurate discussion of error sources, measurement uncertainty, and confidence in your results. An outstanding error discussion is an essential element of a top-notch report.
  • One member of your group should submit a single PDF file to Stellar in advance of the deadline. The filename should consist of the last names of all group members, CamelCased, in alphabetical order, with a .pdf extension. Example: CrickFranklinWatson.pdf.
  • Remember to append your Matlab code at the end of the pdf file.
  • Not including the appendix, the report should not exceed 10 pages.

Part 2 report outline

  1. Haiku:
    • Compose an entertaining, exhilarating, thought-provoking, or melancholy Haiku on the subject of DNA melting.
  2. Abstract:
    • In one paragraph containing six or fewer sentences, summarize the investigation you undertook and key results.
  3. Introduction and Purpose:
    • Provide a succinct introduction to the project, including the purpose of the experiment, relevant background material and/or links to such information.
    • Summarize the ways in which this part of the lab differs from Part 1.
    • Keep the length to one or two short paragraphs, no more than 1/3 of a page.
  4. Apparatus:
    • Document your instrument design.
      • Describe your apparatus with sufficient detail for another person to replicate your work. Assume the reader is familiar with the concepts of 20.309 and has access to course materials.
      • Detail your electronic and optical subsystems. Include component values, gain values, cutoff frequencies, lens focal lengths, and relevant distances.
      • It is not necessary to document construction details, unless you built an instrument that was significantly different than the lab manual suggested.
      • Refer to schematics and diagrams in the lab manual instead of copying them into your report. Use reference designators (such as R7, C2) to refer to call out particular components in schematic diagrams.
    • Why not include a nice snapshot or two of the instrument? So lovely.
  5. Procedure:
    • Document the procedure used to gather your data.
      • Refer to procedures in the lab manual. Describe any changes you made.
      • Report instrument settings for each trial, including control software parameters.
  6. Data:
    • Plot all of your raw data, fluorescence vs. block temperature, on the smallest number of axes that clearly conveys the dataset. Include only data generated by your own group.
      • Data from the many sample runs overlaps, which makes presenting so much data on a small number of axes a real challenge.
      • Devise a combination of line colors, line thicknesses, and marker symbols that produces clear plot. If two sample types have a great deal of overlap, there may be no choice but to plot them on separate axes.
      • One approach that works well for some datasets is to plot a subsampled version of each trial using discrete markers. Vary the color and form to differentiate between sample types and individual trials.
    • Report your signal to noise results.
  7. Analysis:
    • Use bullet points to explain your data analysis methodology.
    • Document the regression model you used to analyze your data
    • Plot $ V_{f,measured} $ and $ V_{f,model} $ versus $ T_{block} $ for a typical run of each samples type. Use the smallest number of axes that clearly conveys the data.
    • For a typical curve, plot residuals versus time, temperature, and fluorescence, (example plot).
    • Provide a table of the best-fit model parameters and confidence intervals for each experimental run. Also include the estimated melting temperature for each run.
    • For at least one experimental trial, plot $ \text{DnaFraction}_{inverse-model} $ versus $ T_{sample} $ (example plot). On the same set of axes plot DnaFraction versus $ T_{sample} $ using the best-fit values of ΔH and ΔS. Finally, plot simulated dsDNA fraction vs. temperature using data from DINAmelt or another melting curve simulator.
  8. Results:
    • Identify your unknown sample (or state that your investigation did not provide a conclusive answer).
    • Quantify the confidence you have in your result.
  9. Discussion:
    • Discuss the validity of assumptions in the regression model.
    • Discuss any atypical results or data you rejected.
    • Compare your data to results from other groups and/or instructor data.
    • Give a bullet point summary of problems you encountered in the lab during part 2 and changes that you made to your instrument and methodology to address those issues.
    • Discuss significant error sources.
      • Consider the entire system: the oligos, dye, the experimental method, and analysis methodology, and any other relevant factors.
      • Indicate whether each source likely caused a systematic or random distortion in the data.
      • Present error sources, error type and their resultant uncertainty on your data and results in a table, if you like.
    • Discuss additional unimplemented changes that might improve your instrument or analysis.

Lab manual sections