16S/18S/ITS Amplicon Sequencing


16S/18S/ITS Amplicon Sequencing

The microbiome has been called another organ or sometimes referred to as our second genome, the amount of microbiota genes is more than 100-fold of human genes, with over 3 million bacterial genes in the gut alone. Over 99% of the bacteria in the gut are anaerobes, many of which have not been studied thoroughly because they cannot be cultured outside of their hosts. The advent of next-generation sequencing has opened new ways to dissect human gut microbial biodiversity. Amplicon sequencing targeted at 16S rRNA, 18S rRNA and ITS, the most commonly sequenced amplicons, allowing for the analysis of genetic variation in specific genomic regions, for the purposes of identifying and analyzing gut microbes including bacteria, archaea, and fungi, in order to understand the dynamics and stability of the human gut microbiota and elucidate its important role in the diagnosis, treatment, and prevention of disease.

Ribosomal RNA is the predominant form of RNA composed around 80% of cellular RNA, which consists of multiple conservative and variable regions, for the benefit of convenient amplification and distinguishable sequences to identify and comparing microbes, with most commonly used fragments distributed within 16S, 18S and ITS.

16S/18S/ITS Amplicon Sequencing

16S rRNA Sequencing: 16S sequencing is widely used for bacterial phylogeny and taxonomy studies, particularly in diverse metagenomics samples. It has also been used to reclassifying bacteria into completely new species or genera, and even describe new species that have never been successfully cultured.

18S rRNA Sequencing: 18S rRNAs are the structural RNAs for the small component of eukaryotic cytoplasmic, which are the homologues of 16S ribosomal RNAs in prokaryotes and mitochondria. Sequence data from 18S rRNA genes are widely used in molecular analysis to reconstruct the evolutionary history of eukaryotes.

ITS Sequencing: Internal transcribed spacer (ITS) is located between the 16S and 23S rRNA genes in bacteria and archaea, while there are two ITSs in eukaryotes genes - ITS1 and ITS2, located each side of 5.8S rRNA gene. ITS1 is the homologue of ITS in bacteria and archaea. ITS is the most widely sequenced DNA region in molecular ecology of fungi, which is typically useful for molecular systematics at the genus and even species level.

Sample Requirements

Region of InterestSample TypeConcentrationMinimum Volume
16SGenomic DNA /
Primary PCR product
1 ng/µl minimum10 µl
18SGenomic DNA /
Primary PCR product
1 ng/µl minimum10 µl
ITSGenomic DNA /
Primary PCR product
1 ng/µl minimum10 µl

Our Advantages

◆ High coverage and highly sensitive
◆ Wide read range
◆ Cost-efficient and fast turnaround time
◆ Bioinformatics support
◆ Specialized in gut microbiota analyses

Creative Proteomics provides 16S/18S/ITS amplicon sequencing services at high depth with high precision to meet the needs of studying population genetics and microbial profiles covering the range of bacteria, archaea, and fungi found in the gastrointestinal tract, in order to research into biodiversity within and interaction between these microbes and human bodies. Our 16S/18S/ITS sequencing service uses oligonucleotide probes designed to target and capture regions of interest – generally hypervariable regions of conserved genes or intergenic regions, followed by next-generation sequencing to meet the goals of efficient genetic variant identification and characterization by comparing against microbial databases and bioinformatic analysis. If you are interested in our amplicon sequencing service and want to find out more, feel free to contact us!

For Research Use Only. Not for use in diagnostic procedures.

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