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Metagenomic Shotgun Sequencing

Relying on advanced sequencing systems and cutting-edge technologies, Creative Proteomics provides global customers with metagenomic shotgun sequencing and analysis services. It helps you to study the structure of microbial populations, the functional activity of genes, the interactions between microorganisms and the relationship between microorganisms and the environment. This technology frees microorganisms from the constraints of isolation and pure culture, expands the scope for the use of microbial resources and provides an effective tool for the study of environmental microbial communities.

Overview

Metagenomic Shotgun Sequencing

Metagenomics is the study of the entire microbial community in a given habitat, using high-throughput sequencing technology to obtain the sum of genomic information of environmental microorganisms and to study the community structure, species classification, phylogeny, gene function and metabolic pathways of environmental microorganisms. Metagenomic sequencing technology has facilitated the exploitation of microbial resources and accelerated the in-depth study of microecological research processes.

Technical advantages of metagenomic shotgun sequencing

  • It is the entire microbial community in a given habitat that is the object of study, without the need for isolation and culture.
  • The experiments do not require PCR and restore the structure of the community relatively objectively, reflecting the true state of microbial survival.
  • A single sequencing can obtain the genetic information of all microorganisms in the environment, and the species information is much richer.
  • Longer gene sequences can be obtained by splicing, with higher resolution of species annotation, and Binning assembly can reconstruct the microbial sketch genome.
  • Sequencing data can be mined in depth from multiple perspectives. In addition to species level studies, genetic and functional level studies can also be carried out, greatly expanding microbial functional metabolism studies.

Applications of metagenomic shotgun sequencing

  • Species classification of microbial communities
  • Functional analysis of microbial communities
  • To identify specific microorganisms in specific environments and discover tolerant strains and associated genes
  • Correlation analysis of gut microbes with disease to reveal microbial differences between diseased and healthy individuals

Service Workflow

For metagenomic shotgun sequencing and analysis, we followed the basic workflow shown below. At the same time, Creative Proteomics has developed a variety of new sample preparation technologies, and we will adjust the experimental methods and optimize the experimental conditions according to the different properties of the samples.

Service Workflow of Metagenomic Shotgun Sequencing -Creative Proteomics

Sample Requirements

  • Soil ≥ 10 g.
  • Faeces = 3-5 g.
  • Blood ≥ 10 mL.
  • Sludge/ sediment = 5-10 g.
  • DNA ≥ 1 μg, concentration ≥ 20 ng/μL, OD260/280 = 1.8 - 2.0.
  • Water samples ≥ 50 L. It is filtered through a 0.22 μm membrane and the membrane is collected.  

Bioinformatics Analysis

Our analysis content is constantly being updated, so if you would like to know more, please contact our technical engineers.

Analysis type Analyze content
Basic analysis

Data quality control and statistics
Sequence assembly
Gene level analysis

  • Gene prediction
  • Non-redundant gene set construction
  • Gene abundance calculation
  • core-pan gene analysis
  • Intergroup gene number analysis
  • Venn diagram (petal plot) analysis
  • Sample correlation analysis
  • Sample hierarchical clustering analysis
Standard bioinformatics analysis

Species level analysis

  • Krona analysis
  • Species composition (Histogram / Heatmap)
  • PCA / PCOA / NMDS analysis
  • Anosim / PERMANOvA / Adonis analysis
  • Metastat species difference analysis
  • LEfSe analysis

Functional level analysis

  • Functional annotation statistics
  • Functional annotation (cOG / KEGG / CAZy / GO / CARD / VFDB)
  • PHI pathogen-host interactions analysis
  • Functional composition (Heatmap)
  • PCA / PCOA / NMDS analysis
  • Anosim / PERMANOVA / Adonis analysis
  • Metastat functional variance analysis
  • LEfSe analysis
Advanced bioinformatics analysis

Correlation and model predictive analytics

  • Species-to-function correspondence analysis
  • Corrplot correlation analysis
  • Correlation network diagram
Personalised bioinformatics analysis Metagenomic binning analysis (contig binning)
Please contact our technical engineers to discuss and evaluate.

Creative Proteomics provides detailed metagenomic shotgun sequencing and analysis services, please contact us for more information. We deserve your trust.

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