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add metro diagram
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.gitattributes

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*.groovy linguist-detectable=false
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*.py linguist-detectable=false
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*.bash linguist-detectable=false
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*.sh linguist-detectable=false
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Dockerfile linguist-detectable=false
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*.mmd linguist-detectable=false

README.md

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### **Overview**
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![Pipeline Overview](figures/metro.svg)
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This pipeline performs context-aware transcript discovery and quantification from long-read single-cell and spatial transcriptomics data. The workflow is divided into three stages:
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**Preprocessing**

figures/logo.png

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figures/metro.mmd

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%%metro title: Bambu-Pipe
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%%metro style: light
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%%metro file: samplesheet_in | CSV | Samplesheet
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%%metro file: ref_genome | FASTA | Reference Genome
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%%metro file: ref_annotation | GTF | Reference Annotation
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%%metro files: in_fastq | FASTQ
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%%metro files: in_bam | BAM
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%%metro file: bam_out | BAM | Output BAM
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%%metro file: se_gene_counts | SE | Gene Counts
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%%metro file: se_unique_counts | SE | Unique Counts
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%%metro file: ext_annotation | GTF | ExtendedAnnotations
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%%metro file: transcript_counts | SE | Transcript Counts
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%%metro line: fastq | FASTQ samples | #f5a623
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%%metro line: bam | Pre-aligned BAM | #0570b0
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%%metro line: em_single | Pseudobulk quantification (single sample) | #9b59b6 | dashed
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%%metro line: em_multi | Pseudobulk quantification (multi-sample) | #6c3483 | dashed
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%%metro line: sc | Single cell quantification | #ff0000 | dashed
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%%metro legend: bl
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%%metro grid: inputs | 0,1
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%%metro grid: preprocessing | 1,1
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%%metro grid: alignment | 2,1
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%%metro grid: bambu | 3,1
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%%metro grid: quantification | 4,1
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graph LR
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subgraph inputs [Inputs]
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%%metro exit: right | fastq, bam
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samplesheet_in[ ]
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ref_genome[ ]
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ref_annotation[ ]
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in_fastq[ ]
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in_bam[ ]
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end
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subgraph preprocessing [FASTQ Preprocessing]
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%%metro entry: left | fastq
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%%metro exit: right | fastq
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chopper_filter[Chopper]
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flexiplex[Flexiplex]
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cutadapt[Cutadapt]
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chopper_filter -->|fastq| flexiplex
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flexiplex -->|fastq| cutadapt
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end
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subgraph alignment [Alignment]
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%%metro entry: left | fastq
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%%metro exit: right | fastq
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minimap_align[Minimap2]
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bam_out[ ]
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minimap_align -->|fastq| bam_out
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end
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subgraph bambu [Transcript Discovery]
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%%metro entry: left | fastq, bam
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%%metro exit: right | sc, em_single, em_multi
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transcript_discovery[Bambu]
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se_gene_counts[ ]
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se_unique_counts[ ]
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ext_annotation[ ]
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transcript_discovery -->|fastq,bam| ext_annotation
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ext_annotation -->|fastq,bam| se_unique_counts
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se_unique_counts -->|fastq,bam| se_gene_counts
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end
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subgraph quantification [Transcript Quantification]
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%%metro entry: left | sc, em_single, em_multi
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seurat_single[Seurat]
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seurat_multi[Seurat + Harmony]
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_cluster_out[ ]
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bambu_em[Bambu]
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transcript_counts[ ]
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seurat_single -->|em_single| _cluster_out
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seurat_multi -->|em_multi| _cluster_out
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_cluster_out -->|em_single,em_multi| bambu_em
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bambu_em -->|sc,em_single,em_multi| transcript_counts
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end
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%% Inter-section edges
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samplesheet_in -->|fastq| chopper_filter
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samplesheet_in -->|bam| transcript_discovery
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ref_genome -->|fastq| chopper_filter
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ref_genome -->|bam| transcript_discovery
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ref_annotation -->|fastq| chopper_filter
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ref_annotation -->|bam| transcript_discovery
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in_fastq -->|fastq| chopper_filter
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in_bam -->|bam| transcript_discovery
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cutadapt -->|fastq| minimap_align
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%% Alignment to Bambu
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bam_out -->|fastq| transcript_discovery
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%% Bambu TD to quantification
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transcript_discovery -->|em_single| seurat_single
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transcript_discovery -->|em_multi| seurat_multi
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transcript_discovery -->|sc| bambu_em

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