Magic c18 aq 3 m resin

1 µg of peptides was injected on a 6600 Sciex TripleTOF mass spectrometer interfaced with an Eksigent NanoLC Ultra 1D Plus system. The peptides were separated on a 75-µm-diameter, 20-cm-long new Objective emitter packed with Magic C18 AQ 3 µm resin (Michrom BioResources) and eluted at 300 nl/min with a linear gradient of 2–30% Buffer B for 60 min (Buffer B: 98% ACN, 0.1% FA). MS data acquisition for the individual meiotic time course samples was performed in data-independent acquisition (DIA) SWATH-MS mode using 64 variable precursor isolation windows with 1-Da overlaps acquired each for 50 ms plus one MS1 scan acquired for 250 ms, as described in Gillet et al. (2012 (link)). Library generation was performed in data-dependent acquisition mode (DDA, top20, with 20 s dynamic exclusion after 1 MS/MS). For either mode, the mass ranges recorded were 360–1,460 m/z for MS1 and 50–2,000 m/z for MS2, and the collision energy was set to 0.0625 × m/z—6.5 with a 15-eV collision energy spread regardless of the precursor charge state.

1 µg of peptides were injected on a 5600 TripleTof mass spectrometer (ABSciex, Concord, Ontario, Canada) interfaced with an Eksigent NanoLC Ultra 1D Plus system (Eksigent, Dublin, CA, USA). The peptides were separated on a 75-µm-diameter, 20-cm long New Objective emitter packed with Magic C18 AQ 3 µm resin (Michrom BioResources) and eluted at 300 nl/min with a linear gradient of 5–35% Buffer A for 120 min (Buffer A: 2% acetonitrile, 0.1% FA; Buffer B: 98% acetonitrile, 0.1% FA). MS data acquisition was performed in either data-dependent acquisition (DDA, top20, with 20 s dynamic exclusion) or data-independent acquisition (DIA) SWATH-MS mode (32 fixed precursor isolation windows of 25 Da width [+1 Da overlap] each acquired for 100 ms plus one MS1 scan acquired for 250 ms) as described in Gillet et al. (2012) (link). The mass ranges recorded were 360–1460 m/z for MS1 and 50–2000 m/z for MS2. For either mode, the collision energy was set to 0.0625 × m/z – 6.5 with a 15 eV collision energy spread regardless of the precursor charge state.