Difference between revisions of "20.109(S17):Complete data analysis (Day6)"
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==Protocols== | ==Protocols== | ||
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+ | ===Part 1: Check alignment of scan image=== | ||
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+ | ===Part 2: Calculate z-scores and p-values=== | ||
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+ | ===Part 3: Determine structures of positive hits=== | ||
==Reagents== | ==Reagents== |
Revision as of 20:58, 31 January 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 a ligand that is bound to the FKBP12 protein. You will analyze the fluorescence emission data collected when your slides were scanned using to 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
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:
Previous day: Scan slides to identify FKBP12 binders