ArticleBioTechniques2019
Triplicate PCR reactions for 16S rRNA gene amplicon sequencing are unnecessary.
Article in BioTechniques, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 35 papers.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
35 citing papers in PubMed.
- HighALPS: ultra-high-throughput marker-gene amplicon library preparation and sequencing on the Illumina NextSeq and NovaSeq Platforms.mSystems · 2026Article
- Effect of perinatal ampicillin or amoxicillin/clavulanate exposure on maternal and infant gut microbiome, metabolome, and infant responses to the 20-valent pneumococcal conjugate vaccine.Gut microbes reports · 2026Article
- Effect of Perinatal Ampicillin or Amoxicillin/Clavulanate Exposure on Maternal and Infant Gut Microbiome, Metabolome, and Infant Responses to the 20-valent Pneumococcal Conjugate Vaccine.bioRxiv : the preprint server for biology · 2025Article
- Role of Vaginal Microbiota and OralBioengineering (Basel, Switzerland) · 2025Article
- Effects of fermented and fiber-rich foods on maternal & offspring microbiome study (FeFiFo-MOMS) - Study design and methods.Contemporary clinical trials · 2025Article
- Biogeographical Distribution of River Microbial Communities in Atlantic Catchments.Environmental microbiology reports · 2025Article
- A Comparison of Three Automated Nucleic Acid Extraction Systems for Human Stool Samples.Microorganisms · 2024Article
- IL-22 resolves MASLD via enterocyte STAT3 restoration of diet-perturbed intestinal homeostasis.Cell metabolism · 2024Article
- Effects of high dose aspartame-based sweetener on the gut microbiota and bone strength in young and aged mice.JBMR plus · 2024Article
- Microbiome-induced Increases and Decreases in Bone Tissue Strength can be Initiated After Skeletal Maturity.bioRxiv : the preprint server for biology · 2024Article
- Effects of long-term high dose aspartame on body mass, bone strength, femoral geometry, and microbiota composition in a young and aged cohort of male and female mice.bioRxiv : the preprint server for biology · 2024Article
- Similarity in milk microbiota in replicates.MicrobiologyOpen · 2023Article
- Optimization of high-throughput 16S rRNA gene amplicon sequencing: an assessment of PCR pooling, mastermix use and contamination.Microbial genomics · 2023Article
- An Environmental DNA Primer for Microbial and Restoration Ecology.Microbial ecology · 2023Review
- A closer look into the microbiome of microalgal cultures.Frontiers in microbiology · 2023Article
- Individuality and stability of the koala (PeerJ · 2023Article
- Best practice for wildlife gut microbiome research: A comprehensive review of methodology for 16S rRNA gene investigations.Frontiers in microbiology · 2023Review
- Gypsum endolithic phototrophs under moderate climate (Southern Sicily): their diversity and pigment composition.Frontiers in microbiology · 2023Article
- A Reproducible and Tunable Synthetic Soil Microbial Community Provides New Insights into Microbial Ecology.mSystems · 2022Article
- Experimental methods modestly impact interpretation of the effect of environmental exposures on the larval zebrafish gut microbiome.Scientific reports · 2022Article
Corrections and comments
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Authors and funding
8 authors.
Funding
Abstract
Conventional wisdom holds that PCR amplification for sequencing should employ pooled replicate reactions to reduce bias due to jackpot effects and chimera formation. However, modern amplicon data analysis employs methods that may be less sensitive to such artifacts. Here we directly compare results from single versus triplicate reactions for 16S amplicon sequencing and find no significant impact of adopting a less labor-intensive single-reaction protocol.
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Registered trials
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.