Amplicon sequencing involves sequencing specific lengths of PCR products or captured fragments, currently primarily using high-throughput sequencing technology to sequence specific genetic material in specific environments. Most natural microorganisms cannot be isolated and cloned using traditional isolation and cultivation methods, posing a significant challenge for qualitative and quantitative analysis of natural microorganisms and exploring microbial diversity. Amplicon sequencing can overcome the weaknesses of traditional isolation and cultivation, allowing for qualitative and quantitative analysis of microorganisms in samples and is widely used in microbial diversity detection.
16S and 18S rDNA are hypervariable regions of the 16s or 18s rDNA genes in bacteria and fungi, while ITS (Internal Transcribed Spacer) is the spacer DNA region between the small and large subunit rRNA genes in bacteria, fungi, and archaea. These specific genetic materials include both evolutionarily conserved and variable regions. Due to the ease of PCR amplification, these can be amplified from small amounts of DNA, and the gene sequences exhibit high variability even among closely related species, making amplicon sequencing widely applicable in taxonomy and molecular systematics.
Amplicon sequencing can be subdivided into the following product types based on different research objectives: 16S rDNA Sequencing:16S rDNA is the gene encoding 16S rRNA and is present in all bacterial genomes. It reflects differences between different genera and can be easily sequenced using sequencing technology. It is currently widely used for the detection and identification of pathogenic bacteria. The variable regions of 16S rRNA are often used for phylogenetic classification of different microbial communities at the genus or species level.
18S rDNA Sequencing:18S rDNA is the DNA sequence encoding the small subunit rRNA in eukaryotic organisms. Structurally, it is divided into conserved and hypervariable regions. The conserved region reflects the phylogenetic relationships between species, while the hypervariable region reflects differences between species. Compared to ITS, 18S rDNA has a more conservative evolutionary rate.
ITS Sequencing:In eukaryotic organisms, the intergenic transcribed spacer between 18S rDNA and 28S rDNA is called the ITS region. Because ITS is a non-coding region, it is under less selective pressure and shows high variability. ITS is moderately conserved and can be used to study taxonomic ranks below the species level.
Amplicon Sequencing Research Targets
Advantages of Amplicon Sequencing Technology: High Identification Efficiency:Compared with traditional identification methods such as cloning or culturing, microbial community 16S/18S/ITS sequencing is a faster and more accurate method;
Dual-region Detection:Flexibility to target one or more variable regions allows for longer sequence reads and more accurate analysis of microbial communities;
Lower Cost:Compared to metagenomic sequencing, it requires less sequencing depth and is more cost-effective;
High Sensitivity:Can identify bacteria with extremely low abundance.
Amplicon Sequencing Experimental Process:
Amplicon Sequencing Experimental Process
Amplicon Sequencing Analysis Process:
Amplicon Sequencing Analysis Process
Advantages of Amplicon Sequencing Service:
Experienced technical personnel can provide a full set of professional services from experimental design, sample testing, to data analysis;
Clear processes reduce unnecessary sample and time wastage, resulting in a short delivery time;
Different extraction methods and library preparation strategies are adopted for samples from different sources to meet the research needs of various environments;
Biotech company BGI has proteomics and metabolomics platforms for multi-omics integrated analysis, enhancing the quality of publications.
Amplicon Sequencing Sample Requirements: Sample Types:Meta samples, such as feces, soil, etc., or Meta DNA samples can be tested. For specific requirements, please consult technical personnel.
