### Input Options | Nextflow parameter name | Type | Description | Help | Default | |--------------------------|------|-------------|------|---------| | fastq | string | FASTQ reads to analyse. | You can provide a single FASTQ, a folder of FASTQs, or a multiplexed folder containing one sub-folder per sample or barcode. | | | bam | string | BAM or uBAM reads to analyse. | You can provide a single BAM or uBAM, a folder of BAMs, or a multiplexed folder containing one sub-folder per sample or barcode. | | | analyse_unclassified | boolean | Include unclassified reads from multiplexed input directories. | | False | | analyse_fail | boolean | Include fail reads from multiplexed input directories. | | False | | fastq_chunk | integer | Maximum number of reads per ingress chunk. | Useful mainly for testing or for splitting very large inputs into smaller pieces. | | ### Reference Options | Nextflow parameter name | Type | Description | Help | Default | |--------------------------|------|-------------|------|---------| | ref_genome | string | Reference genome FASTA. | Required in both discover and fixed_annotation modes. | | | ref_annotation | string | Reference transcript annotation in GTF or GFF format. | Required in both discover and fixed_annotation modes. | | | transcriptome_mode | string | How bambu should prepare the transcriptome model. | Use discover for reference-guided transcript discovery and quantification, or fixed_annotation for quantification only against the supplied annotation. | discover | | direct_rna | boolean | Set this for direct RNA sequencing libraries. | | False | ### Sample Options | Nextflow parameter name | Type | Description | Help | Default | |--------------------------|------|-------------|------|---------| | sample_sheet | string | CSV file describing barcodes, aliases, and optional experimental design columns. | For multiplexed runs, the sample sheet should contain both barcode and alias. For differential analysis it must also contain alias, the condition column, and any extra columns named in `--covariates`. | | | sample | string | Single sample name for singleplexed input or to restrict multiplexed analysis to one sample. | | | ### Analysis Options | Nextflow parameter name | Type | Description | Help | Default | |--------------------------|------|-------------|------|---------| | de_analysis | boolean | Run differential gene expression and differential transcript usage analyses. | | False | | condition_column | string | Main comparison column in the sample sheet. | | condition | | covariates | string | Comma-separated extra sample-sheet columns to adjust for, for example batch. | Each listed name must exist as a column in the sample sheet. | | | reference_level | string | Baseline group for the main comparison column. | If omitted, the workflow will use control when that level exists. | | ### Output Options | Nextflow parameter name | Type | Description | Help | Default | |--------------------------|------|-------------|------|---------| | out_dir | string | Directory for user-facing workflow outputs. | | output | | igv | boolean | Generate an IGV configuration file for the aligned BAM outputs. | | False | ### Advanced Options | Nextflow parameter name | Type | Description | Help | Default | |--------------------------|------|-------------|------|---------| | threads | integer | Thread count to use for the core workflow processes. | | 4 | | mod_codes | string | Comma-separated modified base codes to pass to modkit pileup. | Provide values accepted by `modkit pileup --modified-bases`, for example `A:a,C:m`. If omitted, the workflow infers `primary_base:mod_code` pairs from the BAM with `modkit modbam check-tags`. | | | minimap2_opts | string | Extra command-line options to pass to minimap2. | | | | force_alignment | boolean | Force re-alignment of input BAM files. | Read alignment is skipped if the existing sequence names in the aligned BAM match the provided reference. Enable this if the existing alignments used incorrect minimap2 presets (e.g. missing --splice or direct RNA settings). | False | | ndr | number | Optional bambu novel discovery rate override. | | | | skip_sqanti | boolean | Skip SQANTI3 transcript classification and QC. | | False | | sqanti_skip_orf | boolean | Skip ORF prediction during SQANTI3 QC. | | True | | sqanti_extra_args | string | Extra command-line options to pass to SQANTI3. | | |