The plate reader records values in relative fluorescence units (RFU), which depend on the instrument settings.For each protein (wt and mutant), create plots that overlay the emission spectrum of your protein at each urea concentration, as demonstrated in the following video.These may not be the same concentrations you were planning to use! Be sure to take the urea concentrations from the data file, not from your plan. NOTE: The urea concentrations tested are included in the “Setup” page of the data file.You can also turn on closed captioning for all videos. You may need to view full-screen, and/or manually increase the video quality. NOTE: For all videos, you should be able to actually read the numbers being shown. The following video helps get you oriented to your data and demonstrates how to average your blanks and subtract them from your experimental values.Part 1: Blank and normalize your fluorescence data Manually manipulate the fluorescence data to create two plots:.Fit the data directly in GraphPad Prism to determine:.Blank and normalize your fluorescence data.Your data will be provided by your TAs after you complete the in-lab activities.ĭuring data analysis you will perform these major steps:
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