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• Introduction to CLC Genomics Workbench
  • Contact information
  • Download and installation
    • Program download
    • Installation on Microsoft Windows
    • Installation on macOS
    • 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
  • Plugins
    • Install
    • Uninstall
    • Updating plugins
  • Network configuration
  • CLC Server connection
  • Getting started and latest improvements
• User interface
  • View Area
    • Open view
    • History and Elements 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
    • QIAGEN GeneReader
    • Illumina
    • PacBio
    • Fasta read files
    • Sanger sequencing data
    • Ion Torrent
    • General notes on handling paired data
    • SAM and BAM mapping files
  • Import RNA spike-in controls
  • Import Primers
    • Import Primer Pairs
  • Data export
    • Export formats
    • Export parameters
    • Choosing the exported file name(s)
    • Export in VCF format
    • Export of folders and data elements in CLC format
    • Export of dependent elements
    • Export history
    • Backing up data from the CLC Workbench
    • Export of tables
  • Export graphics to files
    • File formats
  • Export graph data points to a file
  • CLC Server data import and export
  • Copy/paste view output
• Data download
  • Search for Sequences at NCBI
    • NCBI search options
    • Handling of NCBI search results
  • Search for PDB Structures at NCBI
    • Structure search options
    • Handling of NCBI structure search results
    • Save structure search parameters
  • Search for Sequences in UniProt (Swiss-Prot/TrEMBL)
    • 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
• References management
  • Download Genomes
  • QIAGEN Sets
  • Custom Sets
    • Copy to References
    • Export a Custom Data Set
    • Import a Custom Data Set
  • Imported Data
    • Exporting reference data outside of the Reference Data Manager framework
• Running tools, handling results and batching
  • Running tools
  • Handling results
    • Running a tool on a CLC Server
  • Batch processing
    • Standard batch processing
    • Batch overview
    • Parameters for batch runs
    • Running the analysis and organizing the results
• Workflows
  • Creating a workflow
    • Adding workflow elements
    • Configuring workflow inputs
    • Configuring workflow tools
    • 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 Track List
  • Distributing and installing workflows
    • Creating a workflow installation file
    • Installing a workflow
    • Managing workflows
    • Workflow version and update
  • Executing a workflow
  • Open copy of installed workflow
  • Batch launching workflows with multiple inputs
• 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
• Tracks
  • Track types
    • Visualizing, zooming and navigating tracks
    • Showing a track in a table
  • Track lists
    • Adding, removing and reordering tracks
    • 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
  • Merge Annotation Tracks
  • Track Conversion
    • Convert to Tracks
    • Convert from Tracks
  • Annotate and Filter
    • Annotate with Overlap Information
    • Filter Annotations on Name
    • Filter Based on Overlap
  • Graphs
    • Create GC Content Graph
    • Create Mapping Graph
    • Identify Graph Threshold Areas
• Prepare Sequencing Data
  • QC for Sequencing Reads
    • Per-sequence analysis
    • Per-base analysis
    • Over-representation analyses
  • Trim Reads
    • Quality trimming
    • Adapter trimming
    • Trim adapter list
    • Length trimming
    • Trim output
  • Demultiplex Reads
    • An example using Illumina barcoded sequences
• Read mapping
  • Map Reads to Reference
    • Selecting the reads
    • References and masking
    • Mapping parameters
    • Mapping paired reads
    • Non-specific matches
    • Gap placement
    • Mapping computational requirements
    • Reference caching
    • Mapping output options
    • Summary mapping report
  • Reads tracks and stand-alone read mappings
    • Coloring of mapped reads
    • Reads tracks
    • Stand-alone read mapping
  • Local Realignment
    • Method
    • Realignment of unaligned ends
    • Guided realignment
    • Multi-pass local realignment
    • Known limitations
    • Computational requirements
    • Run the Local Realignment tool
  • Merge Read Mappings
  • Remove Duplicate Mapped Reads
    • Algorithm details and parameters
    • Running the duplicate reads removal
  • Extract Consensus Sequence
• Variant detection
  • Variant Detection tools
    • Differences in the variants called by the different tools
    • How the variant detection tools work
    • Detailed information about overlapping paired reads
  • Fixed Ploidy Variant Detection
    • Ploidy and sensitivity
  • Low Frequency Variant Detection
  • Basic Variant Detection
  • Variant Detection - error model estimation
  • Variant Detection - filters
    • General filters
    • Noise filters
  • Variant Detection - the outputs
    • The variant track output
    • The annotated table output
    • The variant detection report
  • Fixed Ploidy and Low Frequency Detection tools: detailed descriptions
    • The Fixed Ploidy Variant Detection tool: Models and methods
    • The Low Frequency Variant Detection tool: Models and methods
  • Variant data
    • Variant tracks
    • The annotated variant table
    • Variant types
  • Copy Number Variant Detection
    • The Copy Number Variant Detection tool
    • Region-level CNV track (Region CNVs)
    • Target-level CNV track (Target CNVs)
    • Gene-level annotation track (Gene CNVs)
    • CNV results report
    • CNV algorithm report
  • Identify Known Mutations from Sample Mappings
    • Run the Identify Known Mutations from Sample Mappings tool
    • Output from the Identify Known Mutations from Sample Mappings tool
  • InDels and Structural Variants
    • Run the InDels and Structural Variants tool
    • The Structural Variants and InDels output
    • The InDels and Structural Variants detection algorithm
    • Theoretically expected structural variant signatures
    • How sequence complexity is calculated
• Resequencing analysis
  • Quality Control for resequencing analyses
    • QC for Targeted Sequencing
    • QC for Read Mapping
    • Whole Genome Coverage Analysis
  • Variant filtering
    • Filter Variants on Custom Criteria
    • Filter against Known Variants
    • Remove Marginal Variants
    • Remove Variants Present in Control Reads
    • Remove Reference Variants
    • Remove Orphan Reference Variants
  • Variant annotation
    • Annotate from Known Variants
    • Remove Information from Variants
    • Annotate with Conservation Score
    • Annotate with Exon Numbers
    • Annotate with Flanking Sequence
  • Variants Comparison
    • Identify Shared Variants
    • Identify Enriched Variants in Case vs Control Samples
    • Trio Analysis
  • Functional consequences
    • Amino Acid Changes
    • Predict Splice Site Effect
    • GO Enrichment Analysis
    • Download 3D Protein Structure Database
    • Link Variants to 3D Protein Structure
• RNA-seq
  • 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
  • PCA for RNA-Seq
    • Principal component analysis plot (2D)
    • Principal component analysis plot (3D)
  • Differential Expression
    • The GLM model
    • Differential Expression in Two Groups
    • Differential Expression for RNA-Seq
    • Output of the Differential Expression tools
  • 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
  • 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
  • The CLC de novo assembly algorithm
    • 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
  • De Novo Assembly
    • Best practices
    • Randomness in the results
    • De novo assembly parameters
    • De novo assembly report
    • De novo assembly output
  • Map Reads to Contigs
• Epigenomics analysis
  • 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
  • Bisulfite Sequencing
    • Detecting DNA methylation
    • Map Bisulfite Reads to Reference
    • Call Methylation Levels
    • Create RRBS-fragment Track
• Utility tools
  • Batch Rename
  • Extract Annotations
  • Sample reads
  • Extract Reads
• Legacy tools
  • Compare Sample Variant Tracks
  • Download Reference Genome Data
  • Identify Differentially Expressed Gene Groups and Pathways
  • Merge Overlapping Pairs
  • Add Information from Overlapping Genes
  • Import Roche 454
  • Create Fold Change Track
  • Add Fold Changes
  • Create Track from Experiment
• Appendix
  • 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
  • Complex variant representations and VCF reference overlap
    • Import of complex variants with reference overlap
  • 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

Prepare Sequencing Data

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Subsections

• QC for Sequencing Reads
• Trim Reads
• Demultiplex Reads

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