Community Analysis Using Next-generation Sequencing

Next-generation sequencing, referred to simply as NGS, is a powerful tool for bacterial detection and microbial community analysis in a high-throughput manner. Creative Biolabs employs 16S rRNA metagenomic sequencing to support high-resolution and low-cost microbial community analysis services.

Introduction of Next-generation Sequencing

NGS allows the rapid sequencing of the base pairs in DNA or RNA samples. The basic sequencing process involves breaking down DNA/RNA into multiple pieces, adding adapters, sequencing libraries, and reassembling them to form a genomic sequence. This technique has a wide variety of applications including whole-genome sequencing, target sequencing, gene expression profiling, chromatin immunoprecipitation sequencing, small RNA sequencing, epigenomic sequencing, metagenomic sequencing, and so on. In microbiome research, 16S rRNA metagenomic sequencing is considered a powerful tool for microbial identification and microbial diversity studies in large sample sets.

Fig.1 The gene/genome-centric approach for the gut microbiome.¹

16S rRNA Sequencing

The prokaryotic 16S rRNA gene with about 1500 bp long is highly conserved between different species of bacteria. It contains nine hypervariable regions (V1-V9) which are commonly used as targets for phylogenetic classification of genus or species in diverse microbial populations. Targeted amplicon sequencing of the 16S rRNA sequencing focus on the analysis of nine hypervariable regions, which improves sequencing coverage, simplifies analysis, and reduces the total sequencing workflow costs. It enables analysis of thousands of microorganisms in one analysis and identification of new strains that may not be found using traditional methods. 16S rRNA sequencing has been reported as an effective method for the characterization of bacterial populations, taxonomical analysis, and species identification.

Workflow

Our Capacities

Creative Biolabs offers NGS-based 16S rRNA analysis services from DNA extraction, preparing genomic and 16S rDNA libraries to sequence. Based on NGS-based 16S rRNA sequencing, we can offer:

• Identification and characterization of hundreds of organisms in one assay
• Determine the relative abundance of microorganisms
• Taxonomic classification
• Community and phylogenetic analysis
• Identifying the bacteria that are difficult to culture

Our Advantages

• More number of sequencing reads: the number of reads reached 100,000, which could detect more sequences and rare microorganisms than Miseq at the same cost.
• Higher data quality
• Higher detection sensitivity: uniform coverage, high detection sensitivity, and higher detection flux enabled more microbial species to be accurately identified.
• Wider coverage: covering a wider range of variable regions, greatly improving the accuracy and sensitivity of subsequent species annotation on sequences.

Sample Requirements

• Samples sources including feces, natural environments, as well as DNA samples
• DNA samples (concentration≥50 ng/μl; volume≥60 μl; purity: OD260/280=1.8~2.0; without degradation and RNA or protein contamination)
• Repeated freezing-thawing should be avoided during the storage of samples
• Transport on dry ice

If you are interested in microbial community analysis using NGS method, please feel free to contact us for more details.

Reference

1. Ji, Boyang, and Jens Nielsen. "From next-generation sequencing to systematic modeling of the gut microbiome." Frontiers in genetics 6 (2015): 219. Distributed under Open Access license CC BY 4.0, without modification.

For Research Use Only. Not intended for use in food manufacturing or medical procedures (diagnostics or therapeutics). Do Not Use in Humans.