Difference between revisions of "20.109(S17):Complete data analysis (Day6)"
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Today is the culminating day of module 1! Hopefully you will identify 'positive hits' from your SMM, or ligands that are able to bind FKBP12. Though you may be able to qualitatively visualize spots that appear to emit more fluorescence, it is important to complete quantitative analysis that supports your observations. | Today is the culminating day of module 1! Hopefully you will identify 'positive hits' from your SMM, or ligands that are able to bind FKBP12. Though you may be able to qualitatively visualize spots that appear to emit more fluorescence, it is important to complete quantitative analysis that supports your observations. | ||
| − | A positive hit denotes a spot on the slide that emits a fluorescence signal significantly higher than the background fluorescence level. In terms of protein binding, a positive hit denotes that the FKBP12 protein is bound to the ligand that was printed at the location of the the fluorescence signal. You will analyze the fluorescence emission data collected | + | A positive hit denotes a spot on the slide that emits a fluorescence signal significantly higher than the background fluorescence level. In terms of protein binding, a positive hit denotes that the FKBP12 protein is bound to the ligand that was printed at the location of the the fluorescence signal. You will analyze the fluorescence emission data collected by the microarray scanner using two quantitative approaches: a z-score and a p-value. Both calculations were explained in detail by Shelby Doyle during the M1D5 lecture and are briefly reviewed here. |
'''z-score''' | '''z-score''' | ||
Revision as of 15:55, 1 February 2017
Contents
Introduction
Today is the culminating day of module 1! Hopefully you will identify 'positive hits' from your SMM, or ligands that are able to bind FKBP12. Though you may be able to qualitatively visualize spots that appear to emit more fluorescence, it is important to complete quantitative analysis that supports your observations.
A positive hit denotes a spot on the slide that emits a fluorescence signal significantly higher than the background fluorescence level. In terms of protein binding, a positive hit denotes that the FKBP12 protein is bound to the ligand that was printed at the location of the the fluorescence signal. You will analyze the fluorescence emission data collected by the microarray scanner using two quantitative approaches: a z-score and a p-value. Both calculations were explained in detail by Shelby Doyle during the M1D5 lecture and are briefly reviewed here.
z-score
p-value
Protocols
Part 1: Check alignment of scan image
Part 2: Calculate z-scores and p-values
Part 3: Determine structures of positive hits
Reagents
Next day: Identify chemical structures common to FKBP12 binders
Previous day: Scan slides to identify FKBP12 binders