DIA-NN 2.1.0 Academia (Data-Independent Acquisition by Neural Networks) Compiled on Mar 25 2025 10:59:54 Current date and time: Sat May 17 23:30:33 2025 CPU: AuthenticAMD AMD Ryzen 9 5950X 16-Core Processor SIMD instructions: AVX AVX2 FMA SSE4.1 SSE4.2 SSE4a Logical CPU cores: 32 diann.exe --f \\YBisei-Server\Bisei-lab\AD_MS-DATA\Ko\2.TrueFDR\250511_Revised\RawFiles\diann_dia_2.1\Mix15-16.wiff.dia --lib \\YBisei-Server\Bisei-lab\AD_MS-DATA\Ko\2.TrueFDR\250511_Revised\250513_diann_2.1_isoform_lib\Mix-8_lib.predicted.speclib --threads 32 --verbose 1 --out \\YBisei-Server\Bisei-lab\AD_MS-DATA\Ko\2.TrueFDR\250511_Revised\250513_SearchResult_diann_2.1_isoform\Mix-8.parquet --qvalue 0.01 --matrices --out-lib \\YBisei-Server\Bisei-lab\AD_MS-DATA\Ko\2.TrueFDR\250511_Revised\250513_SearchResult_diann_2.1_isoform\Mix-8_lib.parquet --gen-spec-lib --reannotate --fasta \\YBisei-Server\Bisei-lab\AD_MS-DATA\Ko\2.TrueFDR\250511_Revised\250511_FASTA_input\clean_Mix-8.fasta --min-pep-len 7 --max-pep-len 30 --min-pr-mz 300 --max-pr-mz 1800 --min-pr-charge 1 --max-pr-charge 4 --cut K*,R* --missed-cleavages 0 --unimod4 --rt-profiling --pg-level 0 --cut --duplicate-proteins Thread number set to 32 Output will be filtered at 0.01 FDR Precursor/protein x samples expression level matrices will be saved along with the main report A spectral library will be generated Library precursors will be reannotated using the FASTA database Min peptide length set to 7 Max peptide length set to 30 Min precursor m/z set to 300 Max precursor m/z set to 1800 Min precursor charge set to 1 Max precursor charge set to 4 In silico digest will involve cuts at K*,R* Maximum number of missed cleavages set to 0 Cysteine carbamidomethylation enabled as a fixed modification The spectral library (if generated) will retain the original spectra but will include empirically-aligned RTs Implicit protein grouping: isoform IDs; this determines which peptides are considered 'proteotypic' and thus affects protein FDR calculation Duplicate proteins in FASTA files will not be skipped DIA-NN will automatically optimise the mass accuracy for the first run of the experiment, use this mode for preliminary analyses only 1 files will be processed [0:00] Loading spectral library \\YBisei-Server\Bisei-lab\AD_MS-DATA\Ko\2.TrueFDR\250511_Revised\250513_diann_2.1_isoform_lib\Mix-8_lib.predicted.speclib [0:02] Library annotated with sequence database(s): \\YBisei-Server\Bisei-lab\AD_MS-DATA\Ko\2.TrueFDR\250511_Revised\250511_FASTA_input\clean_Mix-8.fasta [0:02] Spectral library loaded: 18130 protein isoforms, 21886 protein groups and 1114496 precursors in 353481 elution groups. [0:02] Loading FASTA \\YBisei-Server\Bisei-lab\AD_MS-DATA\Ko\2.TrueFDR\250511_Revised\250511_FASTA_input\clean_Mix-8.fasta [0:07] Reannotating library precursors with information from the FASTA database [0:08] Finding proteotypic peptides (assuming that the list of UniProt ids provided for each peptide is complete) [0:08] 1114496 precursors generated [0:08] Protein names missing for some isoforms [0:08] Gene names missing for some isoforms [0:08] Library contains 0 proteins, and 0 genes [0:08] Initialising library [0:11] File #1/1 [0:11] Loading run \\YBisei-Server\Bisei-lab\AD_MS-DATA\Ko\2.TrueFDR\250511_Revised\RawFiles\diann_dia_2.1\Mix15-16.wiff.dia [0:26] Pre-processing... [0:28] 1704 MS1 and 124392 MS2 scans in 1704 (inferred) and 1704 (encoded) cycles, 981072 precursors in range [0:29] Calibrating with mass accuracies 25 (MS1), 25 (MS2) [1:07] RT window set to 4.9569 [1:07] Peak width: 3.096 [1:07] Scan window radius set to 6 [1:07] Recommended MS1 mass accuracy setting: 31 ppm [3:18] Optimised mass accuracy: 31 ppm [4:03] Main search [5:31] Removing low confidence identifications [5:33] Removing interfering precursors [5:34] Training neural networks on 37044 target and 23371 decoy PSMs [5:50] Number of IDs at 0.01 FDR: 15872 [5:50] Calculating protein q-values [5:50] Number of protein isoforms identified at 1% FDR: 1967 (precursor-level), 1829 (protein-level) (inference performed using proteotypic peptides only) [5:50] Quantification [5:52] Quantification information saved to \\YBisei-Server\Bisei-lab\AD_MS-DATA\Ko\2.TrueFDR\250511_Revised\RawFiles\diann_dia_2.1\Mix15-16.wiff.dia.quant [5:52] Cross-run analysis [5:52] Reading quantification information: 1 files [5:54] Quantifying peptides [5:54] Assembling protein groups [5:54] Quantifying proteins [5:54] Calculating q-values for protein and gene groups [5:54] Calculating global q-values for protein and gene groups [5:54] Protein groups with global q-value <= 0.01: 1879 [5:54] Compressed report saved to \\YBisei-Server\Bisei-lab\AD_MS-DATA\Ko\2.TrueFDR\250511_Revised\250513_SearchResult_diann_2.1_isoform\Mix-8.parquet. Use R 'arrow' or Python 'PyArrow' package to process [5:54] Saving precursor levels matrix [5:54] Precursor levels matrix (1% precursor and protein group FDR) saved to \\YBisei-Server\Bisei-lab\AD_MS-DATA\Ko\2.TrueFDR\250511_Revised\250513_SearchResult_diann_2.1_isoform\Mix-8.pr_matrix.tsv. [5:54] Saving protein group levels matrix [5:54] Protein groups matrix saved to \\YBisei-Server\Bisei-lab\AD_MS-DATA\Ko\2.TrueFDR\250511_Revised\250513_SearchResult_diann_2.1_isoform\Mix-8.pg_matrix.tsv. [5:54] Saving gene group levels matrix [5:54] Gene groups matrix saved to \\YBisei-Server\Bisei-lab\AD_MS-DATA\Ko\2.TrueFDR\250511_Revised\250513_SearchResult_diann_2.1_isoform\Mix-8.gg_matrix.tsv. [5:54] Saving unique genes levels matrix [5:54] Unique genes matrix saved to \\YBisei-Server\Bisei-lab\AD_MS-DATA\Ko\2.TrueFDR\250511_Revised\250513_SearchResult_diann_2.1_isoform\Mix-8.unique_genes_matrix.tsv. [5:55] Manifest saved to \\YBisei-Server\Bisei-lab\AD_MS-DATA\Ko\2.TrueFDR\250511_Revised\250513_SearchResult_diann_2.1_isoform\Mix-8.manifest.txt [5:55] Stats report saved to \\YBisei-Server\Bisei-lab\AD_MS-DATA\Ko\2.TrueFDR\250511_Revised\250513_SearchResult_diann_2.1_isoform\Mix-8.stats.tsv [5:55] Generating spectral library: [5:55] 15871 target and 158 decoy precursors saved [5:55] Spectral library saved to \\YBisei-Server\Bisei-lab\AD_MS-DATA\Ko\2.TrueFDR\250511_Revised\250513_SearchResult_diann_2.1_isoform\Mix-8_lib.parquet