Mari-Lee Odendaal, Mei Ling J N Chu, Kayleigh Arp, Gina J van Beveren, Giske Biesbroek, Justyna Binkowska, Thijs Bosch, James A Groot, Raiza Hasrat, Sjoerd Kuiling, Elske M van Logchem, Paula Lusarreta Parga, Wouter A A de Steenhuijsen Piters, Debby Bogaert
{"title":"Protocol for microbial profiling of low-biomass upper respiratory tract samples.","authors":"Mari-Lee Odendaal, Mei Ling J N Chu, Kayleigh Arp, Gina J van Beveren, Giske Biesbroek, Justyna Binkowska, Thijs Bosch, James A Groot, Raiza Hasrat, Sjoerd Kuiling, Elske M van Logchem, Paula Lusarreta Parga, Wouter A A de Steenhuijsen Piters, Debby Bogaert","doi":"10.1016/j.xpro.2025.103740","DOIUrl":null,"url":null,"abstract":"<p><p>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.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"6 2","pages":"103740"},"PeriodicalIF":1.3000,"publicationDate":"2025-04-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"STAR Protocols","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1016/j.xpro.2025.103740","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0
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.