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Microbiota profiling with NGS technologies is common, with 16S ribosomal RNA gene sequencing. Sequencing the full-length 16S RNA gene is impractical because the majority of NGS platforms produce short reads. The bacterial composition of the habitat under investigation will determine the optimal 16S rRNA hypervariable regions, which can be taxonomically wealthy, but due to variability in profiling results for particular clades, selecting the correct 16S rRNA hypervariable region would vary. The detection of microbes in the urinary tract has been enabled by NGS, and urinary microbiota has also emerged as a hot research field. We obtained urine samples from male volunteers and analyzed their urinary microbiota by sequencing a panel of six amplicons encompassing all nine 16S rRNA hypervariable regions in the world. After systematically comparing their results, we find that V1V2 hypervariable regions help identify the taxa found in urine samples and that V1V2 amplicon sequencing is more appropriate for urinary microbiota profiling.
Source link: https://doi.org/10.1101/2022.01.24.477608
Abstract aims The use of 16S ribosomal RNA gene sequencing analyses has quickly expanded in clinical oral studies. Materials and Methods We obtained 33 supra-interval samples of 11 healthy participants from the biobank. For each participant, we took one sample as baseline and two samples spaced at monthly and quarterly intervals for 16S ribosomal RNA gene sequencing analysis. At any time point, Bray Curtis dissimilarity was present in the cohort. The presence of multiple stable states within an individual's healthy population was shown by high periodicity in multiple stable states. Long-stability Accounting for multi-stability will enhance future research and aid in identifying and classifying the most reliable indicators of diseased, healing, and healthy states. For careful classification of a health-associated group, clinical trials using RNA gene sequencing for comparison should use microbiome specific selection criteria for precise classification of a health-associated group.
Source link: https://doi.org/10.21203/rs.3.rs-1620278/v1
Abstract We investigated the effect of using different sets of 16S RNA primers on bacterial composition, variety, and predicted function in chicken ceca. For DNA isolation, Cecal information from Ross 708 birds aged 1, 3, and 5 weeks old were collected for DNA isolation. Microbiota were distinguished by four distinct groups of bacterial communities according to a unweighted UniFrac distance matrix results. As was previously reported for humans and other animal species, our results show that choosing specific sets of the 16S rRNA primers can have a major effect on microbiota analysis and interpretation of results in chicken.
Source link: https://doi.org/10.1038/s41598-021-91387-w
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