Data-independent acquisition (DIA) mode of mass spectrometry, such as the SWATH-MS technology, enables accurate and consistent measurement of proteins, which is crucial for comparative proteomics studies. However, there is lack of free and easy to implement data analysis protocols that can handle the different data processing steps from raw spectrum files to peptide intensity matrix and its downstream analysis. Here, we provide a data analysis protocol, named diatools, covering all these steps from spectral library building to differential expression analysis of DIA proteomics data. The data analysis tools used in this protocol are open source and the protocol is distributed at Docker Hub as a complete software environment that supports Linux, Windows, and macOS operating systems.
Keywords: DDA; DIA; Data analysis; Mass spectrometry; Proteomics; SWATH-MS; Spectral library.