nf-core/circrna
circRNA quantification, differential expression analysis and miRNA target prediction of RNA-Seq data
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 3 columns, and a header row. See usage docs.
The output directory where the results will be saved. You have to use absolute paths to storage on Cloud infrastructure.
string
Phenotype CSV file specifying the experimental design. If provided, the pipeline will run CIRCTEST.
string
^\S+\.csv$
There are two rules for providing the phenotype CSV file. 1) The 'sample' column must match the sample sheets 'sample' column. 2) The response variable containing the phenotype of primary interest in the experiment must have the column name condition. All other columns included in the file are controlled for in the DESeq2
design.
| sample | condition | replicate |
|----------- |----------- |----------- |
| control_1 | ctr | 1 |
| control_2 | ctr | 2 |
| control_3 | ctr | 3 |
| treated_1 | trt | 1 |
| treated_2 | trt | 2 |
| treated_3 | trt | 3 |
The above phenotype file will identify differentially expressed circRNAs/mRNAs between control and treatment cells, whilst controlling for the effect of variation between replicates: ~ replicates + condition
Path to a CSV file containing BED files that should be used for annotation.
string
^\S+\.csv$
The annotation file should be a CSV file with the following columns: name
, file
and min_overlap
. The name
column should contain a unique identifier for the annotation, the file
column should contain the path to the BED file and the min_overlap
column should contain the minimum overlap required for a circRNA to be considered as overlapping with the annotation. The min_overlap
column is optional and defaults to 0.9 if not provided.
Parameters for circrna discovery.
Comma separated list of circRNA quantification tools to use. Supported tools: ciriquant, circexplorer2, find_circ, circrna_finder, mapsplice, dcc, segemehl
string
circexplorer2
Select one or a combination of circRNA quantification tools for the pipeline e.g:
--tool 'circexplorer2, ciriquant, find_circ'
N.B: Selecting more than one circRNA quantification tool will trigger the circRNA filtering parameter --min_tools
This parameter must be a combination of the following values:ciriquant
, circexplorer2
, find_circ
, circrna_finder
, mapsplice
, dcc
, segemehl
, ciriquant
, circexplorer2
, find_circ
, circrna_finder
, mapsplice
, dcc
, segemehl
Minimum number of reads spanning circRNA back-splice junction required for circRNA to be output by workflow.
integer
1
Filter low confidence circRNAs by removing circRNAs with read counts below a specified value. To disable, set the value to 1 (default).
If both start and end of a pair of BSJs are within max_shift bp, they are considered as the same BSJ.
integer
1
Specify the minimum number of tools circRNAs must be called by to be output by the workflow.
integer
1
When multiple circRNA quantification tools have been provided to --tool
, set a filtering method whereby circRNAs are output if they have been called by at least n quantification tools.
Setting --min_tools
to 1 is the same as taking the union, all circRNAs are included in the output.
Setting --min_tools
to 2 will output circRNAs that have been called by at least 2 quantification tools and so on.
Minimum number of samples a circRNA must be detected in to be output by the workflow.
integer
1
Filter circRNAs by removing circRNAs detected in fewer samples than the specified value. To disable, set the value to 1 (default).
Specify the distance at which the annotation script decides if a candidate is a circRNA or EI-circRNA.
integer
During annotation, if one of the start or end position of a circular candidate imperfectly overlaps an exon boundary, the script will consider positions within 'exon_boundary' (default 0bp) to be an exonic circRNA. If they fall outside of this range, the candidate is assumed to be an exonic-intronic circRNA, and the entire underlying sequence is taken for miRNA analysis, as opposed to just the exonic sequences for canonical exonic circRNAs.
Comma separated list of circRNA quantification tools to use. Supported tools: ciriquant, psirc
string
ciriquant,psirc
^((ciriquant|psirc)(,(ciriquant|psirc))*)+$
Select one or a combination of circRNA quantification tools for the pipeline e.g:
--quantification_tools 'ciriquant,psirc'
Number of bootstrap samples to use during psirc quantification.
integer
30
Define paths and threasholds for miRNA analysis.
path to tab-separated file providing the expression counts of mirnas, which are created in pipeline 'smrnaseq'.
mirna sample1 sample2 sample3 id1 count_sample1 count_sample2 count_sample3 id2 ... ... ...
string
^\S+\.tsv$
Minimum percentage of samples, a miRNA has to be expressed in to pass filtering.
number
0.2
The mirna_min_percentage parameter sets the minimum percentage of samples in which a miRNA must be expressed to pass filtering. The default value is 0.2, which means a miRNA must be detected in at least 20% of the samples to be included in the analysis.
Minimum number of reads, a miRNA is required to have to pass filtering.
integer
5
This parameter determines the minimum number of reads that a miRNA must have to pass filtering. The default is 5, meaning a miRNA must have at least 5 reads across the samples to be considered for analysis.
Specifies the type of correlation to be used when analyzing the relationship between miRNA and transcript expression levels. Valid options are 'pearson' or 'spearman'.
string
pearson
Select the correlation method to be applied in the correlation analysis of miRNAs.
This parameter must be a combination of the following values:pearson
, spearman
Comma separated list of miRNA bindingsite prediction tools to use. Supported tools: miranda, targetscan.
string
miranda,targetscan
^((miranda|targetscan)?,?)*[^,]+$
Select one or a combination of miRNA bindingsite prediction tools for the pipeline e.g:
--mirna_tools 'miranda,targetscan'
Specify the number of votes required for a miRNA to be further considered in downstream analysis.'
integer
1
Controls the number of votes required for a binding site prediction to be considered valid. If a miRNA binding site was predicted by two different tools (e.g., miRanda and TargetScan), it receives two votes. By specifying additional tools for miRNA binding site prediction (using the 'mirna_min_tools' parameter), you can adjust the number of votes required for a binding site to be considered valid.
Parameters used by aligners pertinent to circRNA detection
only used at the genome generation step tells STAR how many bases to concatenate from donor and acceptor sides of the junctions.
integer
100
Minimum overhang for a chimeric junction
integer
10
Minimum overhang for annotated junctions
integer
10
Maximum number of junction to be inserted to the genome on the fly at the mapping stage, including those from annotations and those detected in the 1st step of the 2-pass run
integer
1000000
Minimum length of chimeric segment length. Must be set to a positive value to detect circular junctions.
integer
10
Segment length. Default 25
integer
25
Minimum intron length. Default 20
integer
20
Maximum intron length. Default 1000000
integer
1000000
Minimum alignment length. Default 40
integer
40
Minimum distance between two gapped segments to be considered as fusion candidate. Must set to lower values to be sensitive to circular candidates (e.g 200).
integer
200
Sequencing center information to be added to read group of BAM files.
string
Where possible, save unaligned reads from either STAR, HISAT2 or Salmon to the results directory.
boolean
This may either be in the form of FastQ or BAM files depending on the options available for that particular tool.
Reference genome related files and options required for the workflow.
Save generated reference genome files such as indices, chromosome FASTA files.
boolean
true
Name of iGenomes reference.
string
By using a reference genome build on iGenomes, the gtf, mature, species and index files (bar HISAT2 and segemehl) will be automatically downloaded for you.
See the nf-core website docs for more details.
Path to FASTA genome file.
string
^\S+\.fn?a(sta)?(\.gz)?$
If you use AWS iGenomes, this has already been set for you appropriately.
This parameter is mandatory if --genome
is not specified.
Path to reference GTF file.
string
\.gtf$
This parameter is mandatory if --genome
is not specified. Needs to contain the following attributes: gene_id
, transcript_id
and gene_name
.
Path to FASTA file with mature miRNAs. This parameter needs to be specified to perform miRNA interaction analyses.
string
Typically this will be the mature.fa
file from miRBase. Can be given either as a plain text .fa
file or a compressed .gz
file.
Path to Bowtie index files, surrounded by quotes. No glob pattern required.
string
Path to Bowtie2 index files, surrounded by quotes. No glob pattern required.
string
Path to BWA index directory, surrounded by quotes. No glob pattern required.
string
Path to Hisat2 index directory, surrounded by quotes. No glob pattern required.
string
Minimum memory required to use splice sites and exons in the HiSAT2 index build process.
string
200.GB
^\d+(\.\d+)?\.?\s*(K|M|G|T)?B$
HiSAT2 requires a huge amount of RAM to build a genome index for larger genomes, if including splice sites and exons e.g. the human genome might typically require 200GB. If you specify less than this threshold for the HISAT2_BUILD
process then the splice sites and exons will be ignored, meaning that the process will require a lot less memory. If you are working with a small genome, set this parameter to a lower value to reduce the threshold for skipping this check. If using a larger genome, consider supplying more memory to the HISAT2_BUILD
process.
Path to Segemehl Index file.
string
Path to STAR index directory, surrounded by quotes. No glob pattern required.
string
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
.
Options to adjust read trimming criteria.
Skip the adapter trimming step.
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.
Save the trimmed FastQ files in the results directory.
boolean
By default, trimmed FastQ files will not be saved to the results directory. Specify this flag (or set to true in your config file) to copy these files to the results directory when complete.
Skip FastQC quality control of the sequencing reads.
boolean
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.
Minimum number of trimmed reads below which samples are removed from further processing. Some downstream steps in the pipeline will fail if this threshold is too low.
integer
10000
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
Set the top limit for requested resources for any single job.
Maximum number of CPUs that can be requested for any single job.
integer
50
Use to set an upper-limit for the CPU requirement for each process. Should be an integer e.g. --max_cpus 1
Maximum amount of memory that can be requested for any single job.
string
300.GB
^\d+(\.\d+)?\.?\s*(K|M|G|T)?B$
Use to set an upper-limit for the memory requirement for each process. Should be a string in the format integer-unit e.g. --max_memory '8.GB'
Maximum amount of time that can be requested for any single job.
string
240.h
^(\d+\.?\s*(s|m|h|d|day)\s*)+$
Use to set an upper-limit for the time requirement for each process. Should be a string in the format integer-unit e.g. --max_time '2.h'
Less common options for the pipeline, typically set in a config file.
Display help text.
boolean
Display version and exit.
boolean
Save intermediate files.
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.
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.
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
MultiQC report title. Printed as page header, used for filename if not otherwise specified.
string
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
Show all params when using --help
boolean
By default, parameters set as hidden in the schema are not shown on the command line when a user runs with --help
. Specifying this option will tell the pipeline to show all parameters.
Validation of parameters fails when an unrecognised parameter is found.
boolean
By default, when an unrecognised parameter is found, it returns a warinig.
Validation of parameters in lenient more.
boolean
Allows string values that are parseable as numbers or booleans. For further information see JSONSchema docs.