nf-core/differentialabundance

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.

This file is used to define groups of samples from 'input' to compare. It must contain at least the columns 'variable', 'reference', 'target' and 'blocking', where 'variable' is a column in the input sample sheet, 'reference' and 'target' are values in that column, and blocking is a colon-separated list of additional 'blocking' variables (can be an empty string)

For example an expression matrix output from the nf-core/rnaseq workflow. There must be a column in this matrix for every row in the input sample sheet.

Also used as an identifier in some processes

Currently only 'rnaseq' may be specified. In future options like 'affy_array' are possible.

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.

One feature per row. Note that by default these features will just be stripped from matrices prior to internal processing. To actually use them in e.g. normalisation, set --sizefactors_from_controls

Use supplied control features in normalistion/ scaling operations?

This is used in reporting to refer to the observations. Frequently this is 'sample' (e.g. in RNA-seq experiments), but it may also be desirable to refer to 'pool', or 'individual'.

The variable can be used to define groups and derive a minimum group size upon which to base minimum observation numbers. The rationale for this is to allow retention of features that might be present in only one group. Note that this is consciously NOT filtering with an explicit awareness of groups ("feature must be present in all samples of group A"), since this is known to create biases towards discovery of differential features.

MAD = median absolute deviation. A threshold on this value is used to define observations (samples) as outliers, or not, in exploratory plots. Based on the definition at https://wiki.arrayserver.com/wiki/index.php?title=CorrelationQC.pdf.

Some plots are only generated once, with a single sample grouping, this option defines how that sample grouping is selected. It should be 'auto_pca' (variable selected from the sample sheet with the most association with the first principal component), 'contrasts' (pick the variable associated with the first contrast), or a value specifying a specific column in the observations.

either "Wald" or "LRT", which will then use either Wald significance tests (defined by nbinomWaldTest), or the likelihood ratio test on the difference in deviance between a full and reduced model formula (defined by nbinomLRT)

either "parametric", "local", "mean", or "glmGamPoi" for the type of fitting of dispersions to the mean intensity. See estimateDispersions for description.

either "ratio", "poscounts", or "iterate" for the type of size factor estimation. See estimateSizeFactors for description.

the minimum number of replicates required in order to use replaceOutliers on a sample. If there are samples with so many replicates, the model will be refit after these replacing outliers, flagged by Cook's distance. Set to Inf in order to never replace outliers. It set to Inf for fitType="glmGamPoi".

logical, passed to nbinomWaldTest, default is FALSE, where Wald statistics are assumed to follow a standard Normal

logical, whether independent filtering should be applied automatically

a non-negative value which specifies a log2 fold change threshold. The default value is 0, corresponding to a test that the log2 fold changes are equal to zero. The user can specify the alternative hypothesis using the altHypothesis argument, which defaults to testing for log2 fold changes greater in absolute value than a given threshold. If lfcThreshold is specified, the results are for Wald tests, and LRT p-values will be overwritten.

character which specifies the alternative hypothesis, i.e. those values of log2 fold change which the user is interested in finding. The complement of this set of values is the null hypothesis which will be tested. If the log2 fold change specified by 'name' or by contrast' is written as beta , then the possible values for 'altHypothesis' represent the following alternate hypotheses: 1) greaterAbs: |beta| > lfcThreshold , and p-values are two-tailed 2) lessAbs: |beta| < lfcThreshold , p-values are the maximum of the upper and lower tests. The Wald statistic given is positive, an SE-scaled distance from the closest boundary 3) greater: beta > lfcThreshold 4) less: beta < -lfcThreshold

the method to use for adjusting p-values, see help in R for the p.adjust() function (via ?p.adjust). At time of writing available values were "holm", "hochberg", "hommel", "bonferroni", "BH", "BY", "fdr", "none".

the significance cutoff used for optimizing the independent filtering (by default 0.1). If the adjusted p-value cutoff (FDR) will be a value other than 0.1, alpha should be set to that value.

lower bound on the estimated count (used when calculating contrasts)

'rlog', 'vst' or 'rlog,vst'

'ashr' method is the only method currently implemented

Number of cores to use with DESeq()

logical, whether to blind the transformation to the experimental design

the number of genes to subset to (default 1000)

Select the type of permutation to perform in assessing the statistical significance of the enrichment score. (See 'required fields' at https://www.gsea-msigdb.org/gsea/doc/GSEAUserGuideFrame.html?Run_GSEA_Page for more info)

Specify the number of permutations to perform in assessing the statistical significance of the enrichment score. (See 'required fields' at https://www.gsea-msigdb.org/gsea/doc/GSEAUserGuideFrame.html?Run_GSEA_Page)

See 'basic fields' at https://www.gsea-msigdb.org/gsea/doc/GSEAUserGuideFrame.html?Run_GSEA_Page for a detailed explanation.

See https://www.gsea-msigdb.org/gsea/doc/GSEAUserGuideTEXT.htm#_Metrics_for_Ranking for a detailed explanation.

GSEA ranks the genes in the expression dataset and then analyzes that ranked list of genes. Use this parameter to determine whether to sort the genes using the real (default) or absolute value of the ranking metric.

See 'basic fields' at https://www.gsea-msigdb.org/gsea/doc/GSEAUserGuideFrame.html?Run_GSEA_Page

GSEA ranks the genes in the expression dataset and then analyzes that ranked list of genes. Use this parameter to determine whether to sort the genes in descending (default) or ascending order. Ascending order is usually applicable when the ranking metric is a measure of nearness (how close the genes are to one another) rather than distance.

See 'basic fields' at https://www.gsea-msigdb.org/gsea/doc/GSEAUserGuideFrame.html?Run_GSEA_Page

After filtering from the gene sets any gene not in the expression dataset, gene sets larger than this are excluded from the analysis.

See 'basic fields' at https://www.gsea-msigdb.org/gsea/doc/GSEAUserGuideFrame.html?Run_GSEA_Page

After filtering from the gene sets any gene not in the expression dataset, gene sets smaller than this are excluded from the analysis.

See 'basic fields' at https://www.gsea-msigdb.org/gsea/doc/GSEAUserGuideFrame.html?Run_GSEA_Page

Normalization mode. Method used to normalize the enrichment scores across analyzed gene sets: 'meandiv' (default, GSEA normalizes the enrichment scores as described in Normalized Enrichment Score (NES)) OR 'none' (GSEA does not normalize the enrichment scores).

See 'advanced fields' at https://www.gsea-msigdb.org/gsea/doc/GSEAUserGuideFrame.html?Run_GSEA_Page

Method used to randomly assign phenotype labels to samples for phenotype permutations. Not used for gene_set permutations.

See 'advanced fields' at https://www.gsea-msigdb.org/gsea/doc/GSEAUserGuideFrame.html?Run_GSEA_Page

Set to true (default=false) to use the median of each class, instead of the mean, in the metrics for ranking for genes

See 'advanced fields' at https://www.gsea-msigdb.org/gsea/doc/GSEAUserGuideFrame.html?Run_GSEA_Page.

Number of features (gene or probes) to include in the butterfly plot in the Gene Markers section of the gene set enrichment report.

See 'advanced fields' at https://www.gsea-msigdb.org/gsea/doc/GSEAUserGuideFrame.html?Run_GSEA_Page.

Generates summary plots and detailed analysis results for the top x genes in each phenotype, where x is 20 by default. The top genes are those with the largest normalized enrichment scores.

See 'advanced fields' at https://www.gsea-msigdb.org/gsea/doc/GSEAUserGuideFrame.html?Run_GSEA_Page.

Seed used to generate a random number for phenotype and gene_set permutations: timestamp (default), 149, or user input. The specific seed value (149) generates consistent results, which is useful when testing software.

Set to 'true' (default=false) to save the random ranked lists of genes created by phenotype permutations. When you save random ranked lists, for each permutation, GSEA saves the rank metric score for each gene (the score used to position the gene in the ranked list). Saving random ranked lists is memory intensive; therefore, this parameter is set to false by default.

Set to True (default=false) to create a zip file of the analysis results. The zip file is saved to the output folder with all of the other files generated by the analysis. This is useful for sharing analysis results

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.

"This parameter is mandatory if --genome is not specified."

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.

An email address to send a summary email to when the pipeline is completed - ONLY sent if the pipeline does not exit successfully.

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.