Exercise MSMS2
Topic
Run a target-decoy search to determine false discovery rate
Experimental
Data from a QTof, cysteine residues reduced and alkylated with iodoacetamide (+57).
Search Swiss-Prot with precursor and fragment tolerances of 0.3 Da.
Loose trypsin with 2 missed cleavages. If using Mascot, check the decoy checkbox on the search form.
Data
Right-click this link:
MSMS2.mgf,
and choose Save to file
Tips
- If SwissProt not available, search a mix of mammalian proteins, including human and bovine.
- Make sure you turn off any second pass search. This would discriminate against the decoy, because
decoy peptide matches are scattered randomly across all the database entries
Questions
- How many significant peptide matches can you get at ~ 1% false discovery rate?
- Change the taxonomy filter or database to green plants. What happens to the FDR? Why do you think this is?
Hints |
Mascot results |
Answers
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