Workflow: BD Rhapsody™ WTA Analysis Pipeline

Fetched 2024-11-22 07:58:32 GMT

The BD Rhapsody™ WTA Analysis Pipeline is used to create sequencing libraries from single cell transcriptomes without having to specify a targeted panel. After sequencing, the analysis pipeline takes the FASTQ files, a reference genome file and a transcriptome annotation file for gene alignment. The pipeline generates molecular counts per cell, read counts per cell, metrics, and an alignment file.

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Inputs

ID Type Title Doc
Reads File[] Reads
Subsample Float (Optional) Subsample Reads

Any number of reads >1 or a fraction between 0 < n < 1 to indicate the percentage of reads to subsample.

Tag_Names String[] (Optional) Tag Names

Specify the Sample Tag number followed by - (hyphen) and a sample name to appear in the output files. For example: 4-Ramos. Do not use the special characters: &, (), [], {}, <>, ?, |

Subsample_Tags Float (Optional) Subsample Sample Tags

Any number of reads > 1 or a fraction between 0 < n < 1 to indicate the percentage of tag reads to subsample.

Subsample_seed Integer (Optional) Subsample Seed

For use when replicating a previous subsampling run only. Obtain the seed generated from the log file for the SplitFastQ node.

AbSeq_Reference File[] (Optional) AbSeq Reference
Basic_Algo_Only Boolean (Optional) Disable Refined Putative Cell Calling

Determine putative cells using only the basic algorithm (minimum second derivative along the cumulative reads curve). The refined algorithm attempts to remove false positives and recover false negatives, but may not be ideal for certain complex mixtures of cell types. Does not apply if Exact Cell Count is set.

Exact_Cell_Count Integer (Optional) Exact Cell Count

Set a specific number (>=1) of cells as putative, based on those with the highest error-corrected read count

Reference_Genome File Reference Genome
Sample_Tags_Version https://w3id.org/cwl/view/git/b9ec66d82bc04934fbe9f9088ae7e802d72c4a4e/v1.8/rhapsody_wta_1.8.cwl#main/Sample_Tags_Version/Sample_Tags_Version (Optional) Sample Tags Version

The sample multiplexing kit version. This option should only be set for a multiplexed experiment.

Transcriptome_Annotation File Transcriptome Annotation

Steps

ID Runs Label Doc
Metrics
rhapsody_wta_1.8.cwl#Metrics.cwl (CommandLineTool)
AddtoBam
rhapsody_wta_1.8.cwl#AddtoBam.cwl (CommandLineTool)
IndexBAM
rhapsody_wta_1.8.cwl#IndexBAM.cwl (CommandLineTool)
MergeBAM
rhapsody_wta_1.8.cwl#MergeBAM.cwl (CommandLineTool)
AnnotateR1
rhapsody_wta_1.8.cwl#AnnotateR1.cwl (CommandLineTool)
AnnotateR2
rhapsody_wta_1.8.cwl#AnnotateR2.cwl (CommandLineTool)
BundleLogs
rhapsody_wta_1.8.cwl#BundleLogs.cwl (ExpressionTool)
CheckFastqs
rhapsody_wta_1.8.cwl#CheckFastqs.cwl (CommandLineTool)

CheckFastqs does several quality control routines including: (1) ensuring that read pair file names are formatted correctly and contain a read pair mate; (2) disambiguating the \"Subsample Reads\" input and; (3) if not provided, generating a subsampling seed that the downstream instances can use.

GetDataTable
rhapsody_wta_1.8.cwl#GetDataTable.cwl (CommandLineTool)
AnnotateReads
rhapsody_wta_1.8.cwl#AnnotateReads.cwl (CommandLineTool)
PairReadFiles
rhapsody_wta_1.8.cwl#PairReadFiles.cwl (ExpressionTool)

PairReadsFiles takes an array of split files and pairs them, such that an R1 file is transferred to a QualityFilter with its corresponding R2 file. Each file should be formatted as illumina outputs it from basespace: e.g. sample_L001_R1_001.fastq.gz. After being split, that sample file would be an array files named sample_L001_R1_001-00.fastq, sample_L001_R1_001-01.fastq, etc

QualityFilter
rhapsody_wta_1.8.cwl#QualityFilter.cwl (CommandLineTool)
CheckReference
rhapsody_wta_1.8.cwl#CheckReference.cwl (CommandLineTool)
AnnotateMolecules
rhapsody_wta_1.8.cwl#AnnotateMolecules.cwl (CommandLineTool)
Internal_Settings
rhapsody_wta_1.8.cwl#InternalSettings.cwl (ExpressionTool)
SplitAndSubsample

SplitAndSubsample splits, subsamples and formats read files to be deposited in QualityFilter.

FilteredDataTables
rhapsody_wta_1.8.cwl#FilteredDataTables.cwl (ExpressionTool)
Subsample_Settings
rhapsody_wta_1.8.cwl#SubsampleSettings.cwl (ExpressionTool)
Sparse_to_Dense_File
rhapsody_wta_1.8.cwl#SparsetoDenseFile.cwl (CommandLineTool)
Multiplexing_Settings
rhapsody_wta_1.8.cwl#MultiplexingSettings.cwl (ExpressionTool)
Uncompress_Datatables
Sparse_to_Dense_Datatable
rhapsody_wta_1.8.cwl#SparsetoDense.cwl (CommandLineTool)
Putative_Cell_Calling_Settings
rhapsody_wta_1.8.cwl#PutativeCellSettings.cwl (ExpressionTool)
Sparse_to_Dense_Datatable_Unfiltered
rhapsody_wta_1.8.cwl#SparsetoDense.cwl (CommandLineTool)

Outputs

ID Type Label Doc
Logs Directory Pipeline Logs
Bam_Index File Bam Index
Final_Bam File Final BAM File
Multiplex File[] (Optional)
Data_Tables File[] (Optional) Data Tables
Expression_Data File (Optional) Expression Matrix
Metrics_Summary File Metrics Summary
Cell_Label_Filter File[] (Optional) Cell Label Filter
UMI_Adjusted_Stats File (Optional) UMI Adjusted Statistics
Putative_Cells_Origin File (Optional) Putative Cells Origin
Data_Tables_Unfiltered File[] (Optional) Unfiltered Data Tables
Expression_Data_Unfiltered File (Optional) Unfiltered Expression Matrix
Permalink: https://w3id.org/cwl/view/git/b9ec66d82bc04934fbe9f9088ae7e802d72c4a4e/v1.8/rhapsody_wta_1.8.cwl?part=main