Over 99% of natural microorganisms can not be isolated and cultured clonally, while traditional isolation and culture-dependent methods challenge the study of microorganisms in their natural environment.
Metagenomic studies of genetic material that is directly recovered from environmental samples, have bene tted greatly from advanced NGS technology as a method for the exploration of microbial biodiversity.
16S and 18S rDNA are hypervariable regions in the 16S or 18S rRNA genes in bacteria and fungus, while ITS (Internal Transcribed Spacer) is the spacer DNA between the small-subunit and large-subunit rRNA genes in bacteria, fungi and archaea.
Sequence comparison of 16S/18S/ITS regions is widely used in taxonomy and molecular phylogeny because of the easy ampli cation by PCR, even from low quantities of DNA, while they have a high degree of variation even between closely related species.
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