In 16S rDNA sequencing, high-throughput sequencing technology is used to detect microorganisms (mainly bacteria) in specific environments. Combined with bioinformatics analysis, it provides information on species classification, species abundance, population structure, and system evolution. 16S rDNA is a DNA sequence corresponding to the ribosomal 16S rRNA molecule in the bacterial genome, consisting of 9 hypervariable regions (V) interspersed with conserved regions. For DNAs of different environmental samples, the amplification of the V3-V4 regions of 16S rDNA will help distinguish the majority of bacteria, making it the most commonly used bacterial classification standard.
Advantages
● Ultra-low initial amount of construction, no bias, covering low abundance species
● Good splicing effect, accurate and efficient identification of species
● Independent R&D analytical methods to achieve sample flora composition, group differences and phylogenetic analysis
Bioinformatics Analysis
RiboBio can provide multiple bioinformatics analysis services including bascis and advanced analysis services, welcome to contact order@ribobio.com to know more about it.
FAQs
1. Will the poor quality of the DNAs extracted from samples affect follow-up tests?
Environmental samples, especially the more extreme samples, are difficult to extract DNA, and will normally be slightly degraded and salt-ion contaminated. Successful PCR amplification of V3-V4 is able to support subsequent experiments, and may slightly influence the analysis. If DNA degradation is caused during sample collection, it is recommended to re-prepare the sample.
2. What classification level can be identified by 16S rDNA sequencing?
Since 16S rDNA sequencing is detected by amplifying one or several hypervariable regions, it is generally measured to the “genus” level, and a few to the “species” level. For some bacteria, the sequence similarity in the hypervariable region is very high, or the sequence fragment that distinguishes different bacteria is not in the amplified region, thus making it unable to identify “species”.
3. What is the difference between 16S rDNA and metagenomics?
16S rDNA and metagenomics share many similarities in analysis, such as species classification, abundance analysis, and bacterial comparison. However, the functional prediction accuracy of 16S rDNA is relatively low and can only achieve the third level of KEGG. The metagenomics can carry out gene-level analysis, and analyze the gene function of the different genes, the biological pathways involved and the antibiotic resistance ability thorugh the functional annotation of gene sequences. In the research of environmental microbes, the combination of 16S rDNA and metagenomic sequencing can more accurately study community composition, diversity, evolutionary relationships, and gene functions.
