I have been wondering: what are the similarities and differences between 16S sequencing and 16s metagenomics sequencing? Why can 16S only do function prediction, and the metagenomics can do functional annotation?
In order to find the answer, today CD Genomics shares the esoterica to compare those two sequencing.
16S rDNA sequencing: it refers to the PCR amplification for a certain period in a sample of 16S rDNA by using high-throughput sequencing instruments. It mainly identifies the microbial species and abundance of the bacteria in a specific environment.
16s metagenomics sequencing: it is the sum of all the genetic material in the habitat. With the development of high-throughput sequencing technology, the metagenomics technology has been greatly developed, especially in the application of human intestinal microorganisms. Metagenomic sequencing method avoids the defects of microbial cultivation, greatly broadening the use space of microbial resources, which becomes one of the most important issues in life science research.
16S rdna analysis:
Functional gene annotation and analysis
The way of building
16S rDNA library sequencing process:
DNA quality inspection—primer design and synthesis—PCR amplification and product purification—PCR product quantification and homogenization—Illumina high throughput sequencing
Metagenomic library sequencing process:
DNA quality inspection—DNA fragment—library construction—cluster generation—Illumina high throughput sequencing
16S sequence stitching principle: forward sequencing reads and reverse sequencing reads according to the site of overlap
Stitching software: PANDAseq
The principle of metagenomic stitching: based on the principle of De-Brujin graph by using different sizes of K-mer to assemble the filtered data.
Stitching software: SOAPdenovo
16S species notes: compared to RDF database based on OTU
Metagenomic species notes: comared to reference database of known bacteria, fungi and viruses in NCBI database based on the Clean Reads
The distribution of species composition is different in 16S and metagenomic sequencing.
3.OTU of 16S and the gene of metagenomics
16S is based on the OTU abundance table, the species diversity of Alpha, Beta diversity, the difference between groups and so on.
Metagenomics is based on gene abundance to do CAG analysis, MGS analysis, functional annotation and other analyses.
16S function prediction
metagenomics function analysis
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Original Source: CD Genomics Explores the Difference Between 16S Sequencing and 16s Metagenomics Sequencing