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The GC oven temperature was initially held at 150°C for 2 min, increased to 300°C at 10°C/min and finally held at 300°C for 20 min. Coupled gas chromatography-mass spectrometry (GC-MS) analysis of the samples was conducted also using an HP 6890 gas chromatograph system using the same chromatographic protocol described above.
In the UPLC-ESI-IT-MSn experiments for each parent ion of m/z 1405, m/z 1389, m/z 1239 and m/z 1227 we used the four product ions obtained after MS2, MS3 and MS4 experiments and CID of 30 eV against each ion, using the same chromatographic conditions.
Starting from the confidently identified peptides, an excluding list was made for each sample and the LC-MS/MS analysis was repeated using the same chromatographic conditions and the same acquisition method.
Tetrabromopyrrole (TBP), the characterized inducer, was synthesized according to Palmer [24] and purified from the reaction mixture using the same chromatographic conditions described above for the extract of bacterial strain J010.
Targeted metabolomic analysis was performed using the same chromatographic and spectrometric method as untargeted approach.
Targeted lipidomic analysis was performed using the same chromatographic and spectrometric method as untargeted approach.
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A large number of these proteins have been isolated using basically the same chromatographic strategy, that is, fractionation of the venom by size exclusion chromatography, followed by hydrophobic interaction chromatography of the fractions of interest.
A further attempt to improve the analyses, by analyzing again the same samples using the same parameters and chromatographic conditions, but applying an exclusion list (see Materials and Methods, section 2.6) resulted only in the increase of the percent coverage of some proteins already identified.
Using the same arsenic affinity chromatographic strategy, Menzel et al. identified two arsenic-binding proteins to be tubulin and actin on the basis of their molecular weights.
Two laboratories undertaking the analysis independently tested the curtain flow columns under the same chromatographic conditions, using the same test solutes.
At each measurement wave, physicians' assessments of A1C values (matched to when the questionnaires were completed and using the same high-performance liquid chromatographic assay across sites) served as a criterion measure for glycemic control.
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