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| 1 | +%%metro title: Bambu-Pipe |
| 2 | +%%metro style: light |
| 3 | +%%metro file: samplesheet_in | CSV | Samplesheet |
| 4 | +%%metro file: ref_genome | FASTA | Reference Genome |
| 5 | +%%metro file: ref_annotation | GTF | Reference Annotation |
| 6 | +%%metro files: in_fastq | FASTQ |
| 7 | +%%metro files: in_bam | BAM |
| 8 | +%%metro file: bam_out | BAM | Output BAM |
| 9 | +%%metro file: se_gene_counts | SE | Gene Counts |
| 10 | +%%metro file: se_unique_counts | SE | Unique Counts |
| 11 | +%%metro file: ext_annotation | GTF | ExtendedAnnotations |
| 12 | +%%metro file: transcript_counts | SE | Transcript Counts |
| 13 | +%%metro line: fastq | FASTQ samples | #f5a623 |
| 14 | +%%metro line: bam | Pre-aligned BAM | #0570b0 |
| 15 | +%%metro line: em_single | Pseudobulk quantification (single sample) | #9b59b6 | dashed |
| 16 | +%%metro line: em_multi | Pseudobulk quantification (multi-sample) | #6c3483 | dashed |
| 17 | +%%metro line: sc | Single cell quantification | #ff0000 | dashed |
| 18 | +%%metro legend: bl |
| 19 | +%%metro grid: inputs | 0,1 |
| 20 | +%%metro grid: preprocessing | 1,1 |
| 21 | +%%metro grid: alignment | 2,1 |
| 22 | +%%metro grid: bambu | 3,1 |
| 23 | +%%metro grid: quantification | 4,1 |
| 24 | + |
| 25 | +graph LR |
| 26 | + subgraph inputs [Inputs] |
| 27 | + %%metro exit: right | fastq, bam |
| 28 | + samplesheet_in[ ] |
| 29 | + ref_genome[ ] |
| 30 | + ref_annotation[ ] |
| 31 | + in_fastq[ ] |
| 32 | + in_bam[ ] |
| 33 | + end |
| 34 | + |
| 35 | + subgraph preprocessing [FASTQ Preprocessing] |
| 36 | + %%metro entry: left | fastq |
| 37 | + %%metro exit: right | fastq |
| 38 | + chopper_filter[Chopper] |
| 39 | + flexiplex[Flexiplex] |
| 40 | + cutadapt[Cutadapt] |
| 41 | + chopper_filter -->|fastq| flexiplex |
| 42 | + flexiplex -->|fastq| cutadapt |
| 43 | + end |
| 44 | + |
| 45 | + subgraph alignment [Alignment] |
| 46 | + %%metro entry: left | fastq |
| 47 | + %%metro exit: right | fastq |
| 48 | + minimap_align[Minimap2] |
| 49 | + bam_out[ ] |
| 50 | + minimap_align -->|fastq| bam_out |
| 51 | + end |
| 52 | + |
| 53 | + subgraph bambu [Transcript Discovery] |
| 54 | + %%metro entry: left | fastq, bam |
| 55 | + %%metro exit: right | sc, em_single, em_multi |
| 56 | + transcript_discovery[Bambu] |
| 57 | + se_gene_counts[ ] |
| 58 | + se_unique_counts[ ] |
| 59 | + ext_annotation[ ] |
| 60 | + transcript_discovery -->|fastq,bam| ext_annotation |
| 61 | + ext_annotation -->|fastq,bam| se_unique_counts |
| 62 | + se_unique_counts -->|fastq,bam| se_gene_counts |
| 63 | + end |
| 64 | + |
| 65 | + subgraph quantification [Transcript Quantification] |
| 66 | + %%metro entry: left | sc, em_single, em_multi |
| 67 | + seurat_single[Seurat] |
| 68 | + seurat_multi[Seurat + Harmony] |
| 69 | + _cluster_out[ ] |
| 70 | + bambu_em[Bambu] |
| 71 | + transcript_counts[ ] |
| 72 | + seurat_single -->|em_single| _cluster_out |
| 73 | + seurat_multi -->|em_multi| _cluster_out |
| 74 | + _cluster_out -->|em_single,em_multi| bambu_em |
| 75 | + bambu_em -->|sc,em_single,em_multi| transcript_counts |
| 76 | + end |
| 77 | + |
| 78 | + %% Inter-section edges |
| 79 | + samplesheet_in -->|fastq| chopper_filter |
| 80 | + samplesheet_in -->|bam| transcript_discovery |
| 81 | + |
| 82 | + ref_genome -->|fastq| chopper_filter |
| 83 | + ref_genome -->|bam| transcript_discovery |
| 84 | + |
| 85 | + ref_annotation -->|fastq| chopper_filter |
| 86 | + ref_annotation -->|bam| transcript_discovery |
| 87 | + |
| 88 | + in_fastq -->|fastq| chopper_filter |
| 89 | + in_bam -->|bam| transcript_discovery |
| 90 | + |
| 91 | + cutadapt -->|fastq| minimap_align |
| 92 | + |
| 93 | + %% Alignment to Bambu |
| 94 | + bam_out -->|fastq| transcript_discovery |
| 95 | + |
| 96 | + %% Bambu TD to quantification |
| 97 | + transcript_discovery -->|em_single| seurat_single |
| 98 | + transcript_discovery -->|em_multi| seurat_multi |
| 99 | + transcript_discovery -->|sc| bambu_em |
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