ArticleNucleic acids research2017
Synthetic spike-in standards for high-throughput 16S rRNA gene amplicon sequencing.
Article in Nucleic acids research, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 112 papers, 1 of them a synthesis that pooled it.
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Who cites it
112 citing papers in PubMed, 1 synthesis or guideline pooled it.
- Pooled it
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- Quantitative evaluation of microbiome sequencing resolution under varying experimental conditions using defined mock communities.Scientific reports · 2026Article
- The microbiome across the prostate disease continuum: from health and BPH to prostatitis/CPPS and cancer.Oncogene · 2026Review
- Stable nutritional endosymbiosis across cryptic diversity of a leafhopper species complex.BMC genomics · 2026Article
- Analytic attributes of the 16S rRNA gene sequencing methodology for human gut microbiota characterization.Applied microbiology and biotechnology · 2026Article
- Comparing DNA metabarcoding with light microscopy to identify eukaryotic phytoplankton in the Baltic Sea, Kattegat and Skagerrak.Scientific reports · 2026Article
- Comparison of Bacterial Adhesion on Two Different Suture Materials After Tooth Extraction in Women Receiving Antiresorptive Therapy: An Exploratory Clinical Study with Prospective Data Collection.Journal of clinical medicine · 2026Article
- Accuracy of occurrence and abundance estimates from insect metabarcoding.bioRxiv : the preprint server for biology · 2026Article
- Rhizosphere microbial functions drive ecological stoichiometry in soils across an elevational gradient of a temperate mountain ecosystem.Communications biology · 2026Article
- Changes in the gill and gut microbiota of koi infected with carp edema virus.Veterinary research · 2026Article
- Evaluating preservation effects on honeybee gut microbiota inocula.Microbiology spectrum · 2026Article
- Quantification of the absolute abundance of the jejunal mucosa-associated microbiota of pigs using spike-in control.Microbiome research reports · 2026Article
- EnhancedISME communications · 2026Article
- Converting relative amplicon abundances to absolute abundances via flow cytometry: metagenomic validation and application to long ocean transects.ISME communications · 2026Article
- Validation of a long-read 16S rRNA-gene sequencing approach for analysis of clinical samples and bacterial identification in a routine clinical laboratory.Frontiers in microbiology · 2026Article
52 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
High-throughput sequencing of 16S rRNA gene amplicons (16S-seq) has become a widely deployed method for profiling complex microbial communities but technical pitfalls related to data reliability and quantification remain to be fully addressed. In this work, we have developed and implemented a set of synthetic 16S rRNA genes to serve as universal spike-in standards for 16S-seq experiments. The spike-ins represent full-length 16S rRNA genes containing artificial variable regions with negligible identity to known nucleotide sequences, permitting unambiguous identification of spike-in sequences in 16S-seq read data from any microbiome sample. Using defined mock communities and environmental microbiota, we characterized the performance of the spike-in standards and demonstrated their utility for evaluating data quality on a per-sample basis. Further, we showed that staggered spike-in mixtures added at the point of DNA extraction enable concurrent estimation of absolute microbial abundances suitable for comparative analysis. Results also underscored that template-specific Illumina sequencing artifacts may lead to biases in the perceived abundance of certain taxa. Taken together, the spike-in standards represent a novel bioanalytical tool that can substantially improve 16S-seq-based microbiome studies by enabling comprehensive quality control along with absolute quantification.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.