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6 changes: 3 additions & 3 deletions .nf-core.yml
Original file line number Diff line number Diff line change
Expand Up @@ -14,11 +14,11 @@ nf_core_version: 3.3.2
repository_type: pipeline
template:
author: "Harshil Patel, Phil Ewels, Rickard Hammarén"
description: RNA sequencing analysis pipeline for gene/isoform quantification and
extensive quality control.
description: RNA sequencing analysis pipeline for gene/isoform quantification
and extensive quality control.
force: false
is_nfcore: true
name: rnaseq
org: nf-core
outdir: .
version: 3.20.0dev
version: 3.20.0
2 changes: 1 addition & 1 deletion CHANGELOG.md
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Expand Up @@ -3,7 +3,7 @@
The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/)
and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html).

## 3.20.0dev
## 3.20.0

### Credits

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38 changes: 35 additions & 3 deletions docs/output.md
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Expand Up @@ -215,6 +215,21 @@ When `--remove_ribo_rna` is specified, the pipeline uses [SortMeRNA](https://git
- `star_salmon/`
- `*.Aligned.out.bam`: If `--save_align_intermeds` is specified the original BAM file containing read alignments to the reference genome will be placed in this directory.
- `*.Aligned.toTranscriptome.out.bam`: If `--save_align_intermeds` is specified the original BAM file containing read alignments to the transcriptome will be placed in this directory.
- `salmon.merged.gene_counts.tsv`: Matrix of gene-level raw counts across all samples.
- `salmon.merged.gene_tpm.tsv`: Matrix of gene-level TPM values across all samples.
- `salmon.merged.gene.SummarizedExperiment.rds`: RDS object that can be loaded in R that contains a [SummarizedExperiment](https://bioconductor.org/packages/release/bioc/html/SummarizedExperiment.html) container with the abundance TPM (`tpm`), estimated counts (`counts`) and gene length (`length`), estimated library size-scaled counts (`counts_scaled`), estimated length-scaled counts (`counts_length_scaled`) in the assays slot for genes.
- `salmon.merged.gene_lengths.tsv`: Matrix of average within-sample transcript lengths for each gene across all samples.
- `salmon.merged.gene_counts_scaled.tsv`: Matrix of gene-level library size-scaled estimated counts across all samples.
- `salmon.merged.gene_counts_length_scaled.tsv`: Matrix of gene-level length-scaled estimated counts across all samples.
- `salmon.merged.transcript_counts.tsv`: Matrix of isoform-level raw counts across all samples.
- `salmon.merged.transcript_tpm.tsv`: Matrix of isoform-level TPM values across all samples.
- `tx2gene.tsv`: Tab-delimited file containing gene to transcripts ids mappings.
- `salmon.merged.transcript.SummarizedExperiment.rds`: RDS object that can be loaded in R that contains a [SummarizedExperiment](https://bioconductor.org/packages/release/bioc/html/SummarizedExperiment.html) container with the abundance TPM (`tpm`), estimated isoform-level raw counts (`counts`) and transcript length (`length`) in the assays slot for transcripts.
- `star_salmon/<SAMPLE>/`
- `quant.sf`: Salmon transcript-level quantification results.
- `quant.genes.sf`: Salmon gene-level quantification results.
- `star_salmon/<SAMPLE>/logs/`
- `salmon_quant.log`: Salmon quantification log file.
- `star_salmon/log/`
- `*.SJ.out.tab`: File containing filtered splice junctions detected after mapping the reads.
- `*.Log.final.out`: STAR alignment report containing the mapping results summary.
Expand All @@ -224,6 +239,23 @@ When `--remove_ribo_rna` is specified, the pipeline uses [SortMeRNA](https://git

</details>

:::tip
You can access specific assay matrices from the `SummarizedExperiment` RDS object with the following R code:
:::

```r
library(SummarizedExperiment)

# Load the RDS object
se <- readRDS("salmon.merged.gene.SummarizedExperiment.rds")

# View available assays
assayNames(se)

# Access a specific assay, e.g., length-scaled counts
assay(se, "counts_length_scaled")
```

[STAR](https://github.com/alexdobin/STAR) is a read aligner designed for splice aware mapping typical of RNA sequencing data. STAR stands for *S*pliced *T*ranscripts *A*lignment to a *R*eference, and has been shown to have high accuracy and outperforms other aligners by more than a factor of 50 in mapping speed, but it is memory intensive. Using `--aligner star_salmon` is the default alignment and quantification option.

The STAR section of the MultiQC report shows a bar plot with alignment rates: good samples should have most reads as _Uniquely mapped_ and few _Unmapped_ reads.
Expand Down Expand Up @@ -728,14 +760,14 @@ The principal output files are the same between Salmon and Kallisto:
- `<pseudo_aligner>/`
- `<pseudo_aligner>.merged.gene_counts.tsv`: Matrix of gene-level raw counts across all samples.
- `<pseudo_aligner>.gene_tpm.tsv`: Matrix of gene-level TPM values across all samples.
- `all_samples_gene.SummarizedExperiment.rds`: RDS object that can be loaded in R that contains a [SummarizedExperiment](https://bioconductor.org/packages/release/bioc/html/SummarizedExperiment.html) container with the abundance TPM (`tpm`), estimated counts (`counts`) and gene length (`length`), estimated library size-scaled counts (`counts_scaled`), estimated length-scaled counts (`counts_length_scaled`) in the assays slot for genes.
- `<pseudo_aligner>.merged.gene.SummarizedExperiment.rds`: RDS object that can be loaded in R that contains a [SummarizedExperiment](https://bioconductor.org/packages/release/bioc/html/SummarizedExperiment.html) container with the abundance TPM (`tpm`), estimated counts (`counts`) and gene length (`length`), estimated library size-scaled counts (`counts_scaled`), estimated length-scaled counts (`counts_length_scaled`) in the assays slot for genes.
- `<pseudo_aligner>.merged.gene_lengths.tsv`: Matrix of average within-sample transcript lengths for each gene across all samples.
- `<pseudo_aligner>.merged.gene_counts_scaled.tsv`: Matrix of gene-level library size-scaled estimated counts across all samples.
- `<pseudo_aligner>.merged.gene_counts_length_scaled.tsv`: Matrix of gene-level length-scaled estimated counts across all samples.
- `<pseudo_aligner>.merged.transcript_counts.tsv`: Matrix of isoform-level raw counts across all samples.
- `<pseudo_aligner>.merged.transcript_tpm.tsv`: Matrix of isoform-level TPM values across all samples.
- `tx2gene.tsv`: Tab-delimited file containing gene to transcripts ids mappings.
- `all_samples_transcript.SummarizedExperiment.rds`: RDS object that can be loaded in R that contains a [SummarizedExperiment](https://bioconductor.org/packages/release/bioc/html/SummarizedExperiment.html) container with the abundance TPM (`tpm`), estimated isoform-level raw counts (`counts`) and transcript length (`length`) in the assays slot for transcripts.
- `<pseudo_aligner>.merged.transcript.SummarizedExperiment.rds`: RDS object that can be loaded in R that contains a [SummarizedExperiment](https://bioconductor.org/packages/release/bioc/html/SummarizedExperiment.html) container with the abundance TPM (`tpm`), estimated isoform-level raw counts (`counts`) and transcript length (`length`) in the assays slot for transcripts.

:::tip
You can access specific assay matrices from the `SummarizedExperiment` RDS object with the following R code:
Expand All @@ -745,7 +777,7 @@ You can access specific assay matrices from the `SummarizedExperiment` RDS objec
library(SummarizedExperiment)

# Load the RDS object
se <- readRDS("all_samples_gene.SummarizedExperiment.rds")
se <- readRDS("salmon.merged.gene.SummarizedExperiment.rds") # or kallisto.merged.gene.SummarizedExperiment.rds

# View available assays
assayNames(se)
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2 changes: 1 addition & 1 deletion nextflow.config
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Expand Up @@ -413,7 +413,7 @@ manifest {
mainScript = 'main.nf'
defaultBranch = 'master'
nextflowVersion = '!>=24.10.5'
version = '3.20.0dev'
version = '3.20.0'
doi = 'https://doi.org/10.5281/zenodo.1400710'
}

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