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Database searching settings: tolerance for precursor and fragment masses were 2.0 and 0.5 Da respectively; instrument profile: ESI-Trap; database: MSDB (updated May 15 , 2005; fixed modification: carbamidomethyl (cysteine); variable modification: oxidation (methionine).
In each of the three iterations of assembly, the final stringency settings (tolerance and cut-off) were determined by comparing results of different cut-off and tolerance value combinations.
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MS/MS searches were conducted with the following settings: MS/MS tolerance for precursor and fragment ions between 0.2 and 1 Da depending on the sample, methionine oxidation as variable modification and carbamidomethylation of cysteine as fixed modification.
We could observe that the font size and color of each SSR tag were gradually changed according to different settings of tolerance rate.
Optimisation and position tolerance settings were set at 1.5.
The tolerance settings for peptide identification in Mascot searches were set at 0.05 Da for MS and 0.05 Da for MS/MS.
In these calculations we set (N=2^{9}) and used Matlab's in-built ode45 routine, with standard tolerance settings.
Only the parameters obtained with DegKin Manager deviated slightly, presumably due to suboptimal integration accuracy and convergence tolerance settings.
Typical search criteria used for protein identification included automatic peptide and fragment ion tolerance settings (approximately 10 and 25 ppm, respectively), 1 allowed missed cleavage, fixed carbamidomethyl-cysteine modification and variable methionine oxidation.
PFGE dendrograms were produced with Dice UPGMA with position tolerance settings of 0.5% optimization and 1.0% band position tolerance.
All profiles were compared using the band-matching tool, and uncertain bands were included in the position tolerance settings.
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