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• Introduction
  • The concept of CLC Microbial Genomics Module
  • Contact information
  • System requirements
  • Installing modules
    • Licensing modules
    • Uninstalling modules
  • Installing server extensions
    • Licensing server extensions
• Microbial template workflows
  • Taxonomic Analysis template workflows
    • Data QC and Remove Background Reads
    • Data QC and Taxonomic Profiling
    • Merge and Estimate Alpha and Beta diversities
    • QC, Assemble and Bin Pangenomes
  • Amplicon-Based Analysis template workflows
    • Data QC and OTU Clustering
    • Detect Amplicon Sequence Variants and Assign Taxonomies
    • Estimate Alpha and Beta Diversities
  • Typing and Epidemiology template workflows
    • Compare Variants Across Samples
    • Create MLST Scheme with Sequence Types
    • Map to Specified Reference
    • Type Among Multiple Species
    • Type a Known Species
  • QIAseq Analysis template workflows
    • Analyze QIAseq xHYB Viral Panel Data (Human host)
    • Find QIAseq xHYB AMR Markers (Human host)
  • QIAseq Panel Analysis Assistant
• De Novo Assemble Metagenome
• Amplicon-Based Analysis
  • Normalize OTU Table by Copy Number
  • Filter Samples Based on Number of Reads
  • OTU clustering
    • OTU clustering parameters
    • OTU clustering outputs
    • Importing and exporting OTU abundance tables
  • Remove OTUs with Low Abundance
  • Align OTUs with MUSCLE
  • Detect Amplicon Sequence Variants
    • Detect Amplicon Sequence Variants parameters
    • Detect Amplicon Sequence Variants output
    • Importing ASV abundance tables
  • Classify Long Read Amplicons
    • Classify Long Read Amplicons parameters
    • Classify Long Read Amplicons output
• Taxonomic Analysis
  • Contig Binning
    • Bin Pangenomes by Taxonomy
    • Bin Pangenomes by Sequence
  • Taxonomic Profiling
    • Taxonomic Profiling parameters
    • Taxonomic Profiling output
    • Taxonomic Profiling abundance table
  • Identify Viral Integration Sites
    • The Viral Integration Viewer
    • The Viral Integration Report
• Abundance Analysis
  • Merge Abundance Tables
  • Assign Taxonomies to Sequences in Abundance Table
  • Alpha Diversity
    • Alpha diversity measures
  • Beta Diversity
    • Beta diversity measures
  • PERMANOVA Analysis
  • Differential Abundance Analysis
  • Create Heat Map for Abundance Table
    • Clustering of features and samples
    • The heat map view
    • Create heat map for specific taxonomic level
  • Add Metadata to Abundance Table
• Introduction to Typing and Epidemiology
• Find the best matching reference
  • Find Best Matches using K-mer Spectra
    • From samples best matches to a common reference for all
  • Find Best References using Read Mapping
    • The Find Best References using Read Mapping Report
• Phylogenetic trees using SNPs or k-mers
  • Create SNP Tree
    • SNP tree report
    • SNP tree
    • SNP Matrix
  • Create K-mer Tree
    • Visualization of K-mer Tree for identification of common reference
• MLST Scheme Tools
  • Getting started with the MLST Scheme tools
  • MLST Scheme Visualization and Management
  • Minimum Spanning Trees
    • The Minimum Spanning Tree view
    • Navigating the Tree view
    • The Layout panel
    • The Metadata panel
  • Type With MLST Scheme
    • Type With MLST Scheme results
    • The MLST Typing Result element
  • Add Typing Results to MLST Scheme
  • Identify MLST Scheme from Genomes
• Functional Analysis
  • Find Prokaryotic Genes
  • Annotate with BLAST
  • Annotate with DIAMOND
  • Annotate CDS with Best BLAST Hit
  • Annotate CDS with Best DIAMOND Hit
  • Annotate CDS with Pfam Domains
  • Build Functional Profile
    • Functional profile abundance table
  • Infer Functional Profile
  • Identify Pathways
    • Called Pathways Result
    • The Identified Pathways View
• Drug Resistance Analysis
  • Find Resistance with PointFinder
  • Find Resistance with Nucleotide Database
  • Find Resistance with ShortBRED
    • Resistance abundance table
• Databases for MLST Schemes
  • Create MLST Scheme
  • Download MLST Scheme
  • Import MLST Scheme
• Databases for Amplicon-Based Analysis
  • Download Amplicon-Based Reference Database
• Databases for Taxonomic Analysis
  • Download Curated Microbial Reference Database
    • Extracting a subset of a database
  • Download Custom Microbial Reference Database
    • Database Builder
  • Download Pathogen Reference Database
  • Create Taxonomic Profiling Index
• Databases for Functional Analysis
  • Download Protein Database
  • Download Ontology Database
    • The GO Database View
    • The EC Database View
  • Download Pathway Database
    • The Pathway Database
    • The Pathway View
  • Create DIAMOND Index
  • Import RNAcentral Database
  • Import PICRUSt2 Multiplication Table
• Databases for Drug Resistance Analysis
  • Download Resistance Database
    • ARES Database
• QIAseq 16S/ITS Demultiplexer
• Utility Tools
  • Mask Low-Complexity Regions
    • Mask Low-Complexity Regions Report
  • Result Metadata
    • Create a Result Metadata Table
    • Running an analysis directly from a Result Metadata Table
    • Extend Result Metadata Table
    • Use Genome as Result
• Legacy tools
  • Extract Regions from Tracks
  • Analyze Viral Hybrid Capture Panel Data
• Appendix
  • Using the Assembly ID annotation
• Bibliography

The concept of CLC Microbial Genomics Module

CLC Microbial Genomics Module includes tools for microbial community analysis as well as tools for epidemiological typing of microbial isolates.

Microbiome composition analysis based on 16S rRNA and other commonly used metagenome derived amplicon data is fully supported. The primary output of the clustering, tallying and taxonomic assignment processes is an OTU abundance table that lists the abundances of OTUs in the samples under investigation. In addition, analyses based on whole shotgun metagenomic data are also available, leading to taxonomic profiling abundance tables. CLC Microbial Genomics Module also offers the possibility to investigate biological functions associated with complex communities using Gene Ontology (GO) and Pfam databases to annotate whole shotgun metagenomic data in functional abundance tables. All abundance tables are viewable through a number of intuitive visualization options. Secondary analyses include estimations of alpha and beta diversities, in addition to various statistical tests for differential abundance.

Tools for NGS-MLST typing and identification of antimicrobial resistance genes are included in CLC Microbial Genomics Module to enable epidemiological typing of microbial isolates using NGS data. In cases when the precise identity of the isolated species is not known, the tool automatically detects the most closely related reference genome in NCBI's RefSeq bacterial genome collection and the corresponding MLST scheme from MLST.net or PubMLST.org. The powerful new CLC metadata framework allows fast and intuitive browsing, sorting, filtering and selection of samples and associated metadata, including results obtained during analysis. This metadata framework provides a dashboard-like overview for easy filtering and selection of samples for other analyses such as k-mer or SNP tree reconstruction and visualisation for outbreak analysis.

For convenience, expert-configured workflows for microbiome analysis as well as epidemiological typing allow the user to get from raw NGS reads through data processing and statistical analysis to the final graphical results in very few steps. Reference databases and MLST schemes needed to perform the analyses are automatically downloadable using dedicated tools, and can be easily customized to fit the specific needs of your research.

Unless specified otherwise, all tools described in this manual can be found in the Microbial Genomics Module folder that will be placed in the Toolbox once the plugin is installed.

The CLC Microbial Genomics Module is frequently updated. A detailed list of new features, improvements, bug fixes, and changes is available at https://digitalinsights.qiagen.com/clc-microbial-genomics-module-latest-improvements/.

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