nf-core/rnasplice
rnasplice is a bioinformatics pipeline for RNA-seq alternative splicing analysis
Define where the pipeline should find input data and save output data.
Path to comma-separated file containing information about the samples in the experiment.
string
^\S+\.csv$
You will need to create a design file with information about the samples in your experiment before running the pipeline. Use this parameter to specify its location. It has to be a comma-separated file with 4 columns, and a header row. See usage docs.
Path to comma-separated file containing information about the contrasts in the experiment.
string
Source of input files.
string
The output directory where the results will be saved. You have to use absolute paths to storage on Cloud infrastructure.
string
Email address for completion summary.
string
^([a-zA-Z0-9_\-\.]+)@([a-zA-Z0-9_\-\.]+)\.([a-zA-Z]{2,5})$
Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits. If set in your user config file (~/.nextflow/config
) then you don't need to specify this on the command line for every run.
MultiQC report title. Printed as page header, used for filename if not otherwise specified.
string
Save FastQ files after merging re-sequenced libraries in the results directory.
boolean
Reference genome related files and options required for the workflow.
Name of iGenomes reference.
string
If using a reference genome configured in the pipeline using iGenomes, use this parameter to give the ID for the reference. This is then used to build the full paths for all required reference genome files e.g. --genome GRCh38
.
See the nf-core website docs for more details.
Path to FASTA genome file.
string
^\S+\.fn?a(sta)?(\.gz)?$
This parameter is mandatory if --genome
is not specified. If you don't have a BWA index available this will be generated for you automatically. Combine with --save_reference
to save BWA index for future runs.
Path to GTF annotation file.
string
^\S+\.gtf(\.gz)?$
This parameter is mandatory if --genome
is not specified.
Path to GFF3 annotation file.
string
^\S+\.gff(\.gz)?$
This parameter must be specified if --genome
or --gtf
are not specified.
Path to FASTA transcriptome file.
string
^\S+\.fn?a(sta)?(\.gz)?$
Path to directory or tar.gz archive for pre-built STAR index.
string
Path to directory or tar.gz archive for pre-built Salmon index.
string
Specify if your transcript FASTA file is in GENCODE format.
boolean
If your file is in GENCODE format and you would like to run Salmon i.e. --pseudo_aligner salmon
, you will need to provide this parameter in order to build the Salmon index appropriately.
By default, the pipeline uses the gene_name field to obtain additional gene identifiers from the input GTF file when running Salmon.
string
gene_name
Define the attribute type used to group features in the GTF file when running Salmon.
string
gene_id
If generated by the pipeline save the STAR index in the results directory.
boolean
If an alignment index is generated by the pipeline use this parameter to save it to your results folder. These can then be used for future pipeline runs, reducing processing times.
Directory / URL base for iGenomes references.
string
s3://ngi-igenomes/igenomes/
Do not load the iGenomes reference config.
boolean
Do not load igenomes.config
when running the pipeline. You may choose this option if you observe clashes between custom parameters and those supplied in igenomes.config
.
Parameters used to describe centralised config profiles. These should not be edited.
Git commit id for Institutional configs.
string
master
Base directory for Institutional configs.
string
https://raw.githubusercontent.com/nf-core/configs/master
If you're running offline, Nextflow will not be able to fetch the institutional config files from the internet. If you don't need them, then this is not a problem. If you do need them, you should download the files from the repo and tell Nextflow where to find them with this parameter.
Institutional config name.
string
Institutional config description.
string
Institutional config contact information.
string
Institutional config URL link.
string
Less common options for the pipeline, typically set in a config file.
Display help text.
boolean
Display version and exit.
boolean
Method used to save pipeline results to output directory.
string
The Nextflow publishDir
option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See Nextflow docs for details.
Email address for completion summary, only when pipeline fails.
string
^([a-zA-Z0-9_\-\.]+)@([a-zA-Z0-9_\-\.]+)\.([a-zA-Z]{2,5})$
An email address to send a summary email to when the pipeline is completed - ONLY sent if the pipeline does not exit successfully.
Send plain-text email instead of HTML.
boolean
File size limit when attaching MultiQC reports to summary emails.
string
25.MB
^\d+(\.\d+)?\.?\s*(K|M|G|T)?B$
Do not use coloured log outputs.
boolean
Incoming hook URL for messaging service
string
Incoming hook URL for messaging service. Currently, MS Teams and Slack are supported.
Custom config file to supply to MultiQC.
string
Custom logo file to supply to MultiQC. File name must also be set in the MultiQC config file
string
Custom MultiQC yaml file containing HTML including a methods description.
string
Boolean whether to validate parameters against the schema at runtime
boolean
true
Options to adjust read trimming criteria.
Instructs Trim Galore to remove bp from the 5' end of read 1 (or single-end reads).
integer
Instructs Trim Galore to remove bp from the 5' end of read 2 (paired-end reads only).
integer
Instructs Trim Galore to remove bp from the 3' end of read 1 AFTER adapter/quality trimming has been performed.
integer
Instructs Trim Galore to remove bp from the 3' end of read 2 AFTER adapter/quality trimming has been performed.
integer
Instructs Trim Galore to apply the --nextseq=X option, to trim based on quality after removing poly-G tails.
integer
This enables the option Cutadapt --nextseq-trim=3'CUTOFF
option via Trim Galore, which will set a quality cutoff (that is normally given with -q instead), but qualities of G bases are ignored. This trimming is in common for the NextSeq- and NovaSeq-platforms, where basecalls without any signal are called as high-quality G bases.
Save the trimmed FastQ files in the results directory.
boolean
Use this if your input FastQ files have already been trimmed outside of the workflow or if you're very confident that there is no adapter contamination in your data.
boolean
Skip TrimGalore! FastQC.
boolean
Minimum number of trimmed reads below which samples are flagged in multiqc output.
integer
10000
Defube QC options required for the workflow.
Skip FastQC.
boolean
Skip bigWig file creation.
boolean
true
Options to adjust parameters and filtering criteria for read alignments.
Specifies the alignment algorithm to use - available options are 'star_salmon', or 'star'.
string
Specifies the pseudo aligner to use - available options are 'salmon'. Runs in addition to '--aligner'.
string
Create a CSI index for BAM files instead of the traditional BAI index. This will be required for genomes with larger chromosome sizes.
boolean
When using pre-built STAR indices do not re-extract and use splice junctions from the GTF file.
boolean
Override Salmon library type inferred based on strandedness defined in meta object.
string
See Salmon docs.
Sequencing center information to be added to read group of BAM files.
string
Skip all of the alignment-based processes within the pipeline.
boolean
Where possible, save unaligned reads from either STAR or Salmon to the results directory.
boolean
Save the intermediate BAM files from the alignment step.
boolean
By default, intermediate BAM files will not be saved. The final BAM files created after the appropriate filtering step are always saved to limit storage usage. Set this parameter to also save other intermediate BAM files.
Run Leafcutter annotation workflow.
boolean
Run rMATS workflow.
boolean
true
The cutoff used in the null hypothesis test for differential splicing.
number
0.0001
Use paired statistical model.
boolean
true
The length of each read.
integer
40
Detect splicing events that involve an unannotated splice site.
boolean
Minimum Intron Length.
integer
50
Maximum exon length.
integer
500
Run DEXSeq differential exon usage workflow.
boolean
true
Save pre-processed GFF annotation file.
boolean
Path to GFF3 annotation file.
string
^\S+\.gff(\.gz)?$
Minimum alignment quality required for reads to be counted.
integer
10
Combine overlapping genes into a single aggregate gene.
boolean
true
Save plots of the per gene DEXSeq results.
boolean
true
Plot the N most significant genes from the DEXSeq results.
integer
10
Run edgeR workflow.
boolean
true
Save plots of the per gene edgeR results.
boolean
true
Plot the N most significant genes from the edgeR results.
integer
10
Run DEXSeq differential transcript usage workflow.
boolean
true
Generate estimated counts using dtuScaledTPM or scaledTPM abundance estimates.
string
Minimal number of samples where genes should be expressed.
integer
4
Minimal number of samples where features should be expressed.
integer
2
Minimal proportion of samples where features should be expressed.
integer
2
Minimal gene expression.
integer
10
Minimal feature expression.
integer
10
Minimal proportion for feature expression. This value should be between 0 and 1.
number
0.1
Create sashimi plots using MISO.
boolean
true
List containing identifiers of genes to plot.
string
ENSG00000004961, ENSG00000005302, ENSG00000147403
^[\w-]+(,\s+[\w-]+)*$
New-line separate file containing identifiers of genes to plot.
string
Read length used to calculate coverage.
integer
75
Sashimi figure height (inches).
integer
7
Sashimi figure width (inches).
integer
7
Run SUPPA workflow.
boolean
true
Quantify event inclusion levels (PSIs) from multiple samples.
boolean
true
Quantify isoform inclusion levels (PSIs) from multiple samples.
boolean
true
Expression file containing the abundances of all transcripts (ideally in TPM units).
string
An example can be found at "${projectDir}/assets/tpm.txt"
Redefine genes by clustering together transcripts by genomic stranded overlap and sharing at least one exon.
boolean
true
Space separated list of events to generate.
string
SE SS MX RI FL
Boundary type (only used for local AS events).
string
Variability treshold.
integer
10
Defines the number of nucleotides to display in the output GTF.
integer
100
Minimum total expression of the transcripts involved in the event.
integer
Calculate differential splicing for AS events across multiple conditions with replicates.
boolean
true
Calculate differential splicing for differential transcript usage across multiple conditions with replicates
boolean
true
The method to use to calculate the significance.
string
Integer indicating the number of points in the local area of the delta PSI - average TPM distribution.
integer
1000
Lower-bound for the absolute delta PSI value to test for significance.
integer
Correct the p-values by gene.
boolean
true
Indicates if replicates across conditions are paired.
boolean
true
Family-wise error rate to use for the multiple test correction.
number
0.05
Use the median to calculate the Delta PSI, instead of the mean.
boolean
Minimum expression (calculated as average TPM value within-replicates and between-conditions) to be included in the analysis.
integer
Proportion of samples with nan values allowed per condition to calculate a DeltaPSI .
integer
Cluster events according to PSI values across conditions
boolean
true
Cluster transcripts according to PSI values across conditions
boolean
true
P-value threshold to consider an event significant from the dpsi file.
number
Lower-bound for the absolute delta PSI value to cluster.
number
0.05
Maximum distance (between 0 and 1) to consider two events as members of the same cluster.
number
0.05
Distance metric.
string
Maximum distance in PSI space of an event to a cluster.
integer
Minimum number of events required per cluster.
integer
20
Clustering method to use (DBSCAN, OPTICS).
string
Run isoformswitchanalyzer workflow.
boolean
true
alpha value for differential isoform expression
number
0.05
dIF cutoff value for differential isoform expression
number
0.1