Effects of Host Contamination in 16S rRNA Gene Sequencing of Low Biomass Samples

Effects of Host Contamination in 16S rRNA Gene Sequencing of Low Biomass Samples

Tools

Item Type:	Dataset
Title:	Effects of Host Contamination in 16S rRNA Gene Sequencing of Low Biomass Samples
Abstract:	16S ribosomal RNA (rRNA) gene sequencing is a standard tool for microbial community analysis, yet it presents challenges, especially in low biomass samples where host DNA off-target amplification can occur. Such samples, with limited microbial DNA, amplify host genetic material during PCR, potentially distorting microbial profiles. This issue has been reported in several low-biomass studies. This analysis addresses the distortion of microbial profiles by amplifying host DNA in low-biomass samples. We investigate host contamination in 16S rRNA gene sequencing using a bacterial mock community dilution series spiked with varying amounts of human DNA. Off-target amplification dominated when bacterial mock content was less than 10% compared to human DNA, but total DNA concentration induced minimal bias. We found that post-clustering OTU filtering and reference genome mapping are effective methods for contamination mitigation.
Source:	BioProject
Publisher:	National Center for Biotechnology Information (NCBI)
Date:	28 May 2025
External Fulltext:	View full text on external repository or document server
Related to:	URL URL Type https://edoc.mdc-berlin.de/25988/ Publication https://www.ncbi.nlm.nih.gov/sra?linkname=bioproject_sra_all&from_uid=1268757 External Dataset https://www.ncbi.nlm.nih.gov/bioproject?Db=biosample&DbFrom=bioproject&Cmd=Link&LinkName=bioproject_biosample&LinkReadableName=BioSample&ordinalpos=1&IdsFromResult=1268757 UNSPECIFIED

Repository Staff Only: item control page