Protocol for microbial profiling of low-biomass upper respiratory tract samples

Publication date

2025-06-20

Authors

Odendaal, Mari-Lee
Chu, Mei Ling J N
Arp, Kayleigh
van Beveren, Gina J
Biesbroek, Giske
Binkowska, Justyna
Bosch, Thijs
Groot, James A
Hasrat, Raiza
Kuiling, Sjoerd

Editors

Advisors

Supervisors

Document Type

Article
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License

cc_by

Abstract

The upper respiratory tract (URT) microbiota plays a role in both acute and chronic respiratory health outcomes and consists of multiple ecologically distinct niches, all of which have low bacterial biomass. Here, we present a protocol for microbial profiling of low-biomass URT samples. We describe steps for collecting, storing, and extracting DNA. We then detail procedures for performing 16S rRNA sequencing, using an Illumina MiSeq platform, to characterize microbial communities. For complete details on the use and execution of this protocol, please refer to Odendaal et al.1.

Keywords

Bioinformatics, Health Sciences, Microbiology, Molecular Biology, Sequence analysis, Sequencing, General Neuroscience, General Biochemistry,Genetics and Molecular Biology, General Immunology and Microbiology

Citation

Odendaal, M-L, Chu, M L J N, Arp, K, van Beveren, G J, Biesbroek, G, Binkowska, J, Bosch, T, Groot, J A, Hasrat, R, Kuiling, S, van Logchem, E M, Parga, P L, de Steenhuijsen Piters, W A A & Bogaert, D 2025, 'Protocol for microbial profiling of low-biomass upper respiratory tract samples', STAR Protocols, vol. 6, no. 2, 103740. https://doi.org/10.1016/j.xpro.2025.103740