Hi Skyline team,
we are having issues when trying to export isolation lists for PRMs as multiple scheduled methods. We tried it in Skyline daily (versions 126.96.36.199 & 188.8.131.5295) as well as in Skyline 4.2 but with the same result.
Here is what we are doing and what happens:
- we have a skyline document with DDA MS1 data (196 precursors with according MSMS library), including custom retention time standards, which we would like to use to build a PRM assay
- we have set the time window for scheduling to 5 min resulting in a max of 29 concurrent precursors per window
- we would now like export multiple isolation lists with a max. of 20 concurrent precursors
- when selecting Export Isolation List with the settings "Lumos, multiple methods, ignore proteins, scheduled" I cannot select less than 35 max. concurrent peptides. If I do there is no number of methods calculated and if try to export an error occurs saying: "An error occurred attempting to export. The number of required transitions 35 exceeds the maximum 15.".
What are we doing wrong? Why are 35 transitions required? (BTW, it might be a coincidence but 35 is the value of max. concurrent transitions for a 10 min window.)
Suggestions for solutions would be greatly appreciated!
Many thanks and kind regards,