knitr::opts_chunk$set( collapse = TRUE, comment = "#>" )
A configuration file defines the inputs for the RNA-seq workflow. Update the default parameters to include the Synapse ID where your data is stored and to the factor and continuous variables you want to test in the covariate model selection. The full list of configurable options are:
counts: synID: Required. Synapse ID to counts data frame with identifiers to the metadata as column names and gene ids in a column. version: Optional. Include Synapse file version number (e.g. 3). gene id: Required. Column name that corresponds to the gene ids (e.g. feature). metadata: synID: Required. Synapse ID to cleaned metadata file with sample identifiers in a column and variables of interest as column names. version: Optional. Include Synapse file version number (e.g. 3). sample id: Required. Column name that corresponds to the sample ids (e.g. donorid). biomart: synID: Optional. If left blank, Ensembl will be queried with the gene ids provided in the counts. Otherwise, you may provide the Synapse ID to gene metadata from Ensembl. This must include gene length and GC content in order to implement Conditional Quantile Normalization. version: Optional. Include Synapse file version number (e.g. 3). filters: Required. Column name that corresponds to the gene ids (e.g. ensembl_gene_id). host: Optional. A character vector specifying the BioMart database release version. This specification is highly recommended for a reproducible workflow. Defaults to ensembl.org. organism: Required. A character vector of the organism name. This argument takes partial strings. For example,"hsa" will match "hsapiens_gene_ensembl". exon only: Optional. Set to TRUE if you want gene lengths and GC-content to be calculated only for exons of gene features. Recomended depending on you experimental design paradigm. Default is FALSE, which considers the entire transcript start to stop (ie. includes intronic regions). custom build: Optional. If you want to bulid the biomart object from a user specified or custom GTF and genome FASTA file specify this value as TRUE. Default is FALSE. This would be reccomended for users analyzing data from a model system with a trans-gene inserted into the genome. gtfID: Required IF custom build is set to TRUE. Synapse ID to the user specified GTF file to build the biomart object from. gtfVersion: Optional.Include Synapse file version number (e.g. 3). fastaID: Required IF custom build is set to TRUE. Synapse ID to the user specified genome FASTA file to build the biomart object from. fastaVersion: Optional. Include Synapse file version number (e.g. 3). factors: Required. List of factor variables in brackets. Variables must be present in the metadata as column names (e.g. [ "donorid", "source"]). random_effect: Optional. List of factor variables (must also be included in `factors`) that are to be treated as random effects in the linear regression model. (eg. ["donorid"]) continuous: Required. List of continuous variables in brackets. Variables must be present in the metadata as column names (e.g. [ "rin", "rin2"]). x_var: Required. This is your predictor or primary variable of interest. Additionally, a boxplot will visualize the distribution of continuous variables using the x_var as a dimension. conditions: Optional. Filtering low-expression genes is a common practice to improve sensitivity in detection of differentially expressed genes. Low count genes that have less than a user-defined threshold of counts per million (CPM) in a user-defined percentage of samples per the conditions provided here will be removed. (e.g. ["diagnosis", "sex"]). cpm threshold: Optional. The minium allowable CPM to keep a gene in the analysis. percent threshold:Optional. The percentage of samples that should contain the minimum number of CPM. If a condition is passed, the percentage will apply to the samples in that sub-population. sex check: Optional. The exact variable name that corresponds to reported gender or sex to check the distribution of x and y marker expression across samples. dimensions: Required. Specify the PCA dimensions by variable name. color: shape: size: skip model: Optional. If TRUE, the exploratory data report is run. Model selection is not computed. force null model: Optional. Variables to add to the model aprori eg. sex that users want to account for. force model with: Optional. Force differential expression with this user defined model instead of the output of stepwise regression. cores: Optional. Specify an integer of cores to use with a BiocParallel parallel backend. Null value results in the number of available cores minus one being used. Parallel backend ccurrently only supports BiocParallel::SnowParam(). BatchtoolsParam, MulticoreParam, BiocParallelDoparParam, and SerialParam are not currently supported. de FC: The fold-change (FC) of significant differentially expressed (de) genes must exceed this value. This value will be transformed into log2 FC. de p-value threshold: The adjusted p-value of significant differentially expressed (de) genes must exceed this value. de contrasts: Required. primary: Required. Variable(s) in the metadata to define comparisons between groups. Currently must be either one numeric variable, or one or more catagorical variables. is_numeric_int: Optional. Specifies if there is a numeric interaction variable specified. default (FALSE) numeric: Optional. The numeric in variable which interacts with the primary variable(s). default (NULL) contrasts: Optional. A list specifying contrasts of the primary variable(s) to consider for differential sequencing results if using factor(s) as your primary variable. If not specified all combinations will be tested. If specified this will speed up the pipeline. Specify the contrast with the factor values involved in the contrast seperated by a hyphen. (eg for diagnosis, `contrasts: ["AD-CT"]` where AD is the value in diagnosis column for cases and CT is the value for controls. For multi-level contrasts, eg. `primary: ["diagnosis", "Sex"] would have contrasts specified as; `contrasts: ["ZZ_F-CT_F", "ZZ_M-CT_M"]` to look at cases vs controls in females and cases vs controls in males independently. While the order before or after the hyphen doesn't matter, the order of values before/after the underscore does matter. The value order must be the same as the `primary:` specification. eg. `primary: ["diagnosis","sex"]` must be CT_M while `primary: ["sex","diagnosis"]` must be M_CT. <any named list:> If there are multiple comparisons, set them up as nested lists. visualization gene list: Label this list of genes in the volcano plot. report: Required. The name of your project. This will become the name of your output html file. store output: Required. Folder Synapse Id to store output on Synapse.
Add the following code to your website.
For more information on customizing the embed code, read Embedding Snippets.