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• Introduction to CLC Genomics Workbench
  • Contact information
  • Download and installation
    • Program download
    • Installation on Microsoft Windows
    • Installation on Mac OS X
    • Installation on Linux with an installer
  • System requirements
    • Limitations on maximum number of cores
  • Workbench Licenses
    • Request an evaluation license
    • Download a license using a license order ID
    • Import a license from a file
    • Upgrade license
    • Configure license server connection
    • Download a static license on a non-networked machine
    • Viewing mode
    • Start in safe mode
    • CLC Sequence Viewer vs. Workbenches
  • When the program is installed: Getting started
    • Quick start
  • Plugins
    • Install
    • Uninstall
    • Updating plugins
  • Network configuration
  • Latest improvements
• User interface
  • View Area
    • Open view
    • History and Info views
    • Close views
    • Save changes in a view
    • Undo/Redo
    • Arrange views in View Area
    • Moving a view to a different screen
    • Side Panel
  • Zoom and selection in View Area
    • Zoom in
    • Zoom out
    • Selecting, panning and zooming
  • Toolbox and Status Bar
    • Processes
    • Toolbox
    • Favorites
    • Status Bar
  • Workspace
  • List of shortcuts
• Data management and search
  • Navigation Area
    • Data structure
    • Create new folders
    • Sorting folders
    • Multiselecting elements
    • Moving and copying elements
    • Change element names
    • Delete, restore and remove elements
    • Show folder elements in a table
  • Metadata
    • Importing Metadata
    • Advanced Metadata Import
    • Associating data elements with metadata
    • Working with data and metadata
  • Working with tables
    • Filtering tables
  • Customized attributes on data locations
    • Filling in values
    • What happens when a clc object is copied to another data location?
    • Searching
  • Local search
    • Quick search
    • Advanced search
• User preferences and settings
  • General preferences
  • View preferences
    • Import and export Side Panel settings
  • Data preferences
  • Advanced preferences
  • Export/import of preferences
  • View settings for the Side Panel
• Printing
  • Selecting which part of the view to print
  • Page setup
  • Print preview
• Import/export of data and graphics
  • Standard import
    • External files
  • Import tracks
    • GFF3 format
  • Import high-throughput sequencing data
    • Illumina
    • PacBio
    • Fasta read files
    • Sanger sequencing data
    • Ion Torrent
    • Complete Genomics
    • General notes on handling paired data
    • SAM and BAM mapping files
  • Import RNA spike-in controls
  • Data export
    • Export of folders and multiple elements in CLC format
    • Export of dependent elements
    • Export history
    • The CLC format
    • Backing up data from the CLC Workbench
    • Export of tables
  • Export graphics to files
    • File formats
  • Export graph data points to a file
  • Copy/paste view output
• Data download
  • Download reference genome data
    • Selecting data types for download
    • Cytogenetic ideograms
  • Search for Sequences at NCBI
    • NCBI search options
    • Handling of NCBI search results
  • Search for structures at NCBI
    • Structure search options
    • Handling of NCBI structure search results
    • Save structure search parameters
  • UniProt (Swiss-Prot/TrEMBL) search
    • UniProt search options
    • Handling of UniProt search results
    • Save UniProt search parameters
  • SRA search
    • SRA search options
    • SRA search output
    • Downloading reads and metadata from SRA
    • How reads are downloaded
  • Sequence web info
• Running tools, handling results and batching
  • Running tools
  • Handling results
  • Batch processing
    • Standard batch processing
    • Batch overview
    • Parameters for batch runs
    • Running the analysis and organizing the results
    • Batch launching workflows with multiple inputs
• Workflows
  • Creating a workflow
    • Adding workflow elements
    • Configuring workflow elements
    • Locking and unlocking parameters
    • Connecting workflow elements
    • Output
    • Input
    • Layout
    • Input modifying tools
    • Workflow validation
    • Workflow creation helper tools
    • Adding to workflows
    • Snippets in workflows
    • Change the order of tracks in the Genome Browser View
  • Distributing and installing workflows
    • Creating a workflow installation file
    • Installing a workflow
    • Managing workflows
    • Workflow identification and versioning
    • Automatic update of workflow elements
  • Executing a workflow
  • Open copy of installed workflow
• Viewing and editing sequences
  • View sequence
    • Sequence settings in Side Panel
    • Selecting parts of the sequence
    • Editing the sequence
    • Sequence region types
  • Circular DNA
    • Using split views to see details of the circular molecule
    • Mark molecule as circular and specify starting point
  • Working with annotations
    • Viewing annotations
    • Adding annotations
    • Edit annotations
    • Removing annotations
  • Element information
  • View as text
  • Sequence Lists
• BLAST search
  • Running BLAST searches
    • BLAST at NCBI
    • BLAST against local data
  • Output from BLAST searches
    • Graphical overview for each query sequence
    • Overview BLAST table
    • BLAST graphics
    • BLAST HSP table
    • BLAST hit table
    • Extracting a consensus sequence from a BLAST result
  • Local BLAST databases
    • Make pre-formatted BLAST databases available
    • Download NCBI pre-formatted BLAST databases
    • Create local BLAST databases
  • Manage BLAST databases
  • Bioinformatics explained: BLAST
    • How does BLAST work?
    • Which BLAST program should I use?
    • Which BLAST options should I change?
    • Explanation of the BLAST output
    • I want to BLAST against my own sequence database, is this possible?
    • What you cannot get out of BLAST
    • Other useful resources
• 3D Molecule Viewer
  • Importing molecule structure files
    • From the Protein Data Bank
    • From your own file system
    • BLAST search against the PDB database
    • Import issues
  • Viewing molecular structures in 3D
    • Updating old structure files
  • Customizing the visualization
    • Visualization styles and colors
    • Project settings
  • Tools for linking sequence and structure
    • Show sequence associated with molecule
    • Link sequence or sequence alignment to structure
    • Transfer annotations between sequence and structure
  • Protein structure alignment
    • The Align Protein Structure dialog box
    • Example: alignment of calmodulin
    • The Align Protein Structure algorithm
• General sequence analyses
  • Extract Annotations
  • Extract sequences
  • Shuffle sequence
  • Dot plots
    • Create dot plots
    • View dot plots
    • Bioinformatics explained: Dot plots
    • Bioinformatics explained: Scoring matrices
  • Local complexity plot
  • Sequence statistics
    • Bioinformatics explained: Protein statistics
  • Join sequences
  • Pattern discovery
    • Pattern discovery search parameters
    • Pattern search output
  • Motif Search
    • Dynamic motifs
    • Motif search from the Toolbox
    • Java regular expressions
  • Create motif list
• Nucleotide analyses
  • Convert DNA to RNA
  • Convert RNA to DNA
  • Reverse complements of sequences
  • Reverse sequence
  • Translation of DNA or RNA to protein
  • Find open reading frames
    • Open reading frame parameters
• Protein analyses
  • Protein charge
  • Antigenicity
  • Hydrophobicity
    • Hydrophobicity graphs along sequence
    • Bioinformatics explained: Protein hydrophobicity
  • Pfam domain search
    • Download of Pfam database
    • Running Pfam Domain Search
  • Secondary structure prediction
  • Protein report
  • Reverse translation from protein into DNA
    • Bioinformatics explained: Reverse translation
  • Proteolytic cleavage detection
    • Bioinformatics explained: Proteolytic cleavage
• Primers
  • Primer design - an introduction
    • General concept
    • Scoring primers
  • Setting parameters for primers and probes
    • Primer Parameters
  • Graphical display of primer information
    • Compact information mode
    • Detailed information mode
  • Output from primer design
  • Standard PCR
    • When a single primer region is defined
    • When both forward and reverse regions are defined
    • Standard PCR output table
  • Nested PCR
  • TaqMan
  • Sequencing primers
  • Alignment-based primer and probe design
    • Specific options for alignment-based primer and probe design
    • Alignment based design of PCR primers
    • Alignment-based TaqMan probe design
  • Analyze primer properties
  • Find binding sites and create fragments
    • Binding parameters
    • Results - binding sites and fragments
  • Order primers
• Sequencing data analyses
  • Importing and viewing trace data
    • Trace settings in the Side Panel
  • Trim sequences
    • Trimming using the Trim tool
    • Manual trimming
  • Assemble sequences
  • Assemble sequences to reference
  • Sort sequences by name
  • Add sequences to an existing contig
  • View and edit contigs and read mappings
    • View settings in the Side Panel
    • Editing a contig or read mapping
    • Sorting reads
    • Read conflicts
    • Using the mapping
    • Extract parts of a mapping
    • Variance table
  • Reassemble contig
  • Secondary peak calling
• Cutting and cloning
  • Restriction site analyses
    • Dynamic restriction sites
    • Restriction Site Analysis
  • Restriction enzyme lists
  • Molecular cloning
    • Introduction to the cloning editor
    • The cloning workflow
    • Manual cloning
    • Insert restriction site
  • Gateway cloning
    • Add attB sites
    • Create entry clones (BP)
    • Create expression clones (LR)
  • Gel electrophoresis
    • Gel view
• Sequence alignment
  • Create an alignment
    • Gap costs
    • Fast or accurate alignment algorithm
    • Aligning alignments
    • Fixpoints
  • View alignments
    • Bioinformatics explained: Sequence logo
  • Edit alignments
    • Realignment
  • Join alignments
  • Pairwise comparison
    • The pairwise comparison table
    • Bioinformatics explained: Multiple alignments
• Phylogenetic trees
  • K-mer Based Tree Construction
  • Create tree
  • Model Testing
  • Maximum Likelihood Phylogeny
    • Bioinformatics explained
  • Tree Settings
    • Minimap
    • Tree layout
    • Node settings
    • Label settings
    • Background settings
    • Branch layout
    • Bootstrap settings
    • Visualizing metadata
    • Node right click menu
  • Metadata and phylogenetic trees
    • Table Settings and Filtering
    • Add or modify metadata on a tree
    • Undefined metadata values on a tree
    • Selection of specific nodes
• RNA structure
  • RNA secondary structure prediction
    • Selecting sequences for prediction
    • Secondary structure prediction parameters
    • Structure as annotation
  • View and edit secondary structures
    • Graphical view and editing of secondary structure
    • Tabular view of structures and energy contributions
    • Symbolic representation in sequence view
    • Probability-based coloring
  • Evaluate structure hypothesis
    • Selecting sequences for evaluation
    • Probabilities
  • Structure scanning plot
    • Selecting sequences for scanning
    • The structure scanning result
  • Bioinformatics explained: RNA structure prediction by minimum free energy minimization
    • The algorithm
    • Structure elements and their energy contribution
• Trimming, multiplexing and sequencing quality control
  • Trim Reads
    • Quality trimming
    • Adapter trimming
    • Trim adapter list
    • Length trimming
    • Trim output
  • Demultiplex reads
    • An example using Illumina barcoded sequences
  • Sequencing data quality control
    • QC Sequencing Report Content
    • Adapters
    • Running the quality control tool
  • Merge overlapping pairs
    • Using quality scores when merging
    • Report of merged pairs
• Tracks
  • Track lists
    • Zooming and navigating track views
    • Adding, removing and reordering tracks
    • Showing a track in a table
    • Open track from a track list in table view
    • Finding annotations on the genome
    • Extract sequences from tracks
    • Creating track lists in workflows
  • Retrieving reference data tracks
  • Merging tracks
  • Converting data to tracks and back
    • Convert to tracks
    • Convert from tracks
  • Annotate and filter tracks
    • Annotate with overlap information
    • Extract reads based on overlap
    • Filter annotations on name
    • Filter Based on Overlap
  • Graphs
    • Create GC Content Graph
    • Create Mapping Graph
    • Identify Graph Threshold Areas
• Read mapping
  • Map Reads to Reference
    • Selecting reads and reference
    • Including or excluding regions (masking)
    • Mapping parameters
    • Mapping paired reads
    • Non-specific matches
    • Gap placement
    • Mapping computational requirements
    • Reference caching
  • Mapping output
    • Mapping output options
    • Mapped reads coloring
    • Reads track output from a read mapping
    • Stand-alone read mapping
    • Mapping table
    • Mapping view settings
    • Overlapping paired reads
    • Find broken pair mates
  • Mapping reports
    • Summary mapping report
    • Detailed mapping report
  • Mapping SOLid reads in color space
    • Viewing color space information
    • Mapping in color space
  • Local realignment
    • Method
    • Realignment of unaligned ends
    • Guided realignment
    • Multi-pass local realignment
    • Known limitations
    • Computational requirements
    • How to run the Local Realignment tool
  • Merge mapping results
  • Remove duplicate mapped reads
    • Algorithm details and parameters
    • Running the duplicate reads removal
  • Extract consensus sequence
  • Sample reads
• Resequencing
  • Create Statistics for Target Regions
    • Running the Create Statistics for Target Regions
    • Coverage summary report
    • Per-region statistics
    • Coverage table
    • Coverage graph
  • InDels and Structural Variants
    • How to run the InDels and Structural Variants tool
    • The Structural Variants and InDels output
    • The InDels and Structural Variants detection algorithm
    • The InDels and Structural Variants detection algorithm - Step 1: Creating Left- and Right breakpoint signatures
    • The InDels and Structural Variants detection algorithm - Step 2: Creating Structural variant signatures
    • Theoretically expected structural variant signatures
    • How sequence complexity is calculated
  • Coverage analysis
  • Variant Detectors - overview
    • Differences in the variants called by the different tools
    • How the variant detection tools work
  • Fixed Ploidy Variant Detection
    • Ploidy and sensitivity
  • Low Frequency Variant Detection
  • Basic Variant Detection
  • Variant Detectors - error model estimation
  • Variant Detectors - filters
    • General filters
    • Noise filters
  • Variant Detectors - the outputs
    • The variant track output
    • The annotated table output
    • The report
  • The Fixed Ploidy and Low Frequency variant callers: detailed descriptions
    • The Fixed Ploidy Variant Caller: Models and methods
    • The Low Frequency Variant caller: Models and methods
  • Variant data
    • Variant tracks
    • The annotated variant table
    • Variant types
  • Detailed information about overlapping paired reads
  • Annotate and filter variants
    • Filter against known variants
    • Annotating from known variants
    • Annotate with exon numbers
    • Annotate with flanking sequence
    • Identify candidate variants
    • Filter marginal variant calls
    • Filter reference variants
  • Comparing variants
    • Compare variants within group
    • Compare sample variants
    • Fisher exact test
    • Trio analysis
    • Filter Against Control Reads
  • Predicting functional consequences
    • Amino acid changes
    • Predict splice site effect
    • GO enrichment analysis
    • Conservation score annotation
    • Link Variants to 3D Protein Structure
    • Download 3D Protein Structure Database
  • Identify Known Mutations from Sample Mappings
    • How to run the Identify Known Mutations from Sample Mappings tool
    • Output from the Identify Known Mutations from Sample Mappings tool
• RNA-Seq Analysis tools
  • RNA-Seq analysis
    • Specifying reads and reference
    • Defining mapping options for RNA-Seq
    • The EM estimation algorithm
    • Calculating expression values from RNA-Seq
    • Specifying RNA-Seq outputs
    • Expression tracks
    • Reads track
    • RNA-Seq report
    • Gene fusion reporting
  • Create Combined RNA-Seq Report
  • Advanced RNA-Seq Tools
    • TMM Normalization
    • Metadata for RNA-Seq
  • PCA for RNA-Seq
    • Principal component analysis plot (2D)
    • Principal component analysis plot (3D)
  • Differential Expression for RNA-Seq
    • The statistical model
    • Output of the Differential Expression for RNA-Seq tool
    • Statistical comparison tracks
    • The volcano plot
  • Create Heat Map for RNA-Seq
    • Clustering of features and samples
    • The heat map view
  • Create Expression Browser
    • The expression browser
  • Create Venn Diagram for RNA-Seq
    • Venn diagram table view
  • Gene Set Test
• Microarray and Small RNA Analysis
  • Small RNA analysis
    • Extract and count
    • Downloading miRBase
    • Annotating and merging small RNA samples
    • Working with the small RNA sample
    • Exploring novel miRNAs
  • Experimental design
    • Setting up an experiment
    • Organization of the experiment table
    • Adding annotations to an experiment
    • Scatter plot view of an experiment
    • Cross-view selections
  • Working with tracks and experiments
    • Data structures for transcriptomics
    • Running the Create Track from Experiment tool
    • Interpreting the results of the Create Track from Experiment tool
    • Visualizing RNA-Seq read tracks for the experiment
  • Transformation and normalization
    • Selecting transformed and normalized values for analysis
    • Transformation
    • Normalization
  • Quality control
    • Creating box plots - analyzing distributions
    • Hierarchical clustering of samples
    • Principal component analysis
  • Statistical analysis - identifying differential expression
    • Empirical analysis of DGE
    • Tests on proportions
    • Gaussian-based tests
    • Corrected p-values
    • Volcano plots - inspecting the result of the statistical analysis
  • Feature clustering
    • Hierarchical clustering of features
    • K-means/medoids clustering
  • Annotation tests
    • Hypergeometric tests on annotations
    • Gene set enrichment analysis
  • General plots
    • Histogram
    • MA plot
    • Scatter plot
• De novo sequencing
  • De novo assembly
    • Best practices
    • How it works
    • Resolve repeats using reads
    • Automatic paired distance estimation
    • Optimization of the graph using paired reads
    • AGP export
    • Bubble resolution
    • Converting the graph to contig sequences
    • Summary
    • Randomness in the results
    • SOLiD data support in de novo assembly
    • De novo assembly parameters
    • De novo assembly report
  • Map Reads to Contigs
• Epigenomics
  • ChIP-Seq Analysis
    • Quality Control of ChIP-Seq data
    • Learning peak shapes
    • Applying peak shape filters to call peaks
    • Running the Transcription Factor ChIP-Seq tool
  • Annotate with nearby gene information
• Legacy tools
  • Import Roche 454
  • Import SOLiD
• Appendix
  • Use of multi-core computers
  • Graph preferences
  • BLAST databases
    • Peptide sequence databases
    • Nucleotide sequence databases
    • Adding more databases
  • Proteolytic cleavage enzymes
  • Restriction enzymes database configuration
  • Technical information about modifying Gateway cloning sites
  • IUPAC codes for amino acids
  • IUPAC codes for nucleotides
  • Formats for import and export
    • List of bioinformatic data formats
    • List of graphics data formats
  • SAM/BAM export format specification
    • Flags
  • Gene expression annotation files and microarray data formats
    • GEO (Gene Expression Omnibus)
    • Affymetrix GeneChip
    • Illumina BeadChip
    • Gene ontology annotation files
    • Generic expression and annotation data file formats
  • Translation Tables
    • 1. Standard
    • 2. Vertebrate Mitochondrial
    • 3. Yeast Mitochondrial
    • 4. Mold Mitochondrial; Protozoan Mitochondrial; Coelenterate Mitochondrial; Mycoplasma; Spiroplasma
    • 5. Invertebrate Mitochondrial
    • 6. Ciliate Nuclear; Dasycladacean Nuclear; Hexamita Nuclear
    • 9. Echinoderm Mitochondrial; Flatworm Mitochondrial
    • 10. Euplotid Nuclear
    • 11. Bacterial and Plant Plastid
    • 12. Alternative Yeast Nuclear
    • 13. Ascidian Mitochondrial
    • 14. Alternative Flatworm Mitochondrial
    • 15. Blepharisma Macronuclear
    • 16. Chlorophycean Mitochondrial
    • 21. Trematode Mitochondrial
    • 22. Scenedesmus Obliquus Mitochondrial
    • 23. Thraustochytrium Mitochondrial
    • 24. Pterobranchia Mitochondrial
    • 25. Candidate Division SR1 and Gracilibacteria
  • Custom codon frequency tables
  • Comparison of track comparison tools
  • Matrices for alignment calculation
    • PAM30 log-odds matrix
    • PAM60 log-odds matrix
    • BLOSUM42 log-odds matrix
    • BLOSUM62 log-odds matrix
    • BLOSUM80 log-odds matrix
• Bibliography

System requirements

• Windows 7, Windows 8, Windows 10 or Windows Server 2012
• Mac OS X 10.9, 10.10, 10.11 and 10.12
• Linux: RHEL 6.0 and later, SUSE 13.1 and later. Minimum linux versions needed for the SRA download functionality: Red Hat 6, Fedora 12, and SUSE 11.
• 64 bit operating system
• 2 GB RAM required
• 4 GB RAM recommended
• 1024 x 768 display required
• 1600 x 1200 display recommended
• Intel or AMD CPU required

• Special requirements for the 3D Molecule Viewer System requirements
  • A graphics card capable of supporting OpenGL 2.0.
  • Updated graphics drivers. Please make sure the latest driver for the graphics card is installed .

  System Recommendations

  • A discrete graphics card from either Nvidia or AMD/ATI. Modern integrated graphics cards (such as the Intel HD Graphics series) may also be used, but these are usually slower than the discrete cards.

  Indirect rendering (such as x11 forwarding through ssh), remote desktop connection/VNC, and running in virtual machines is not supported.

• Special requirements for read mapping. The numbers below give minimum and recommended memory for systems running mapping and analysis tasks. The requirements suggested are based on the genome size.
  • E. coli K12 (4.6 megabases)
    • Minimum: 2 GB RAM
    • Recommended: 4 GB RAM
  • C. elegans (100 megabases) and Arabidopsis thaliana (120 megabases)
    • Minimum: 2 GB RAM
    • Recommended: 4 GB RAM
  • Zebrafish (1.5 gigabases)
    • Minimum: 2 GB RAM
    • Recommended: 4 GB RAM
  • Human (3.2 gigabases) and Mouse (2.7 gigabases)
    • Minimum: 6 GB RAM
    • Recommended: 8 GB RAM

• Special requirements for de novo assembly. De novo assembly may need more memory than stated above - this depends both on the number of reads, error profile and the complexity and size of the genome. See http://resources.qiagenbioinformatics.com/white-papers/White_paper_on_de_novo_assembly_4.pdf for examples of the memory usage of various data sets.

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Subsections

• Limitations on maximum number of cores

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