ArticlePloS one2019
Miniaturization and optimization of 384-well compatible RNA sequencing library preparation.
Article in PloS one, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 38 papers.
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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
38 citing papers in PubMed.
- Impact of doxycycline post-exposure prophylaxis for sexually transmitted infections on the gut microbiome and antimicrobial resistome.Nature medicine · 2025Trial
- Integrated host-microbe plasma metagenomics for sepsis diagnosis in a prospective cohort of critically ill adults.Nature microbiology · 2022Trial
- Metronidazole treatment rapidly reduces genital inflammation through effects on bacterial vaginosis-associated bacteria rather than lactobacilli.The Journal of clinical investigation · 2022Trial
- Characterization of recombinase-based genetic parts and circuits using nanopore sequencing.Nature communications · 2026Article
- Upper and Lower Respiratory Tract Compartmentalization in Pediatric Stem Cell Transplantation.American journal of respiratory cell and molecular biology · 2026Article
- Host-Microbe Multiomic Profiling Predicts Mortality in Sepsis.American journal of respiratory and critical care medicine · 2026Article
- Upper and Lower Respiratory Tract Compartmentalization in Pediatric Stem Cell Transplantation.medRxiv : the preprint server for health sciences · 2025Article
- Infection in Childhood Arterial Ischemic Stroke: Metagenomic Next-Generation Sequencing Results of the VIPS II Study.Stroke · 2025Article
- Epigenetic attenuation of interferon signaling is associated with aging-related improvements in systemic lupus erythematosus.Science translational medicine · 2025Article
- Pathobiological signatures of dysbiotic lung injury in pediatric patients undergoing stem cell transplantation.Nature medicine · 2024Article
- Vaginal Lactobacillus crispatus persistence following application of a live biotherapeutic product: colonization phenotypes and genital immune impact.Microbiome · 2024Article
- Article
- Doxycycline post-exposure prophylaxis for sexually transmitted infections impacts the gut antimicrobial resistome.Research square · 2024Article
- Article
- Pulmonary microbiome and transcriptome signatures reveal distinct pathobiologic states associated with mortality in two cohorts of pediatric stem cell transplant patients.medRxiv : the preprint server for health sciences · 2023Article
- Affordable, accurate and unbiased RNA sequencing by manual library miniaturization: A case study in barley.Plant biotechnology journal · 2023Article
- Metagenomic sequencing of post-mortem tissue samples for the identification of pathogens associated with neonatal deaths.Nature communications · 2023Article
- Maximizing the potential of high-throughput next-generation sequencing through precise normalization based on read count distribution.mSystems · 2023Article
- Treatment Success Following Standard Antibiotic Treatment for Bacterial Vaginosis Is Not Associated With Pretreatment Genital Immune or Microbial Parameters.Open forum infectious diseases · 2023Article
- Evaluation of miniaturized Illumina DNA preparation protocols for SARS-CoV-2 whole genome sequencing.PloS one · 2023Article
Corrections and comments
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Authors and funding
5 authors.
Funding
Abstract
Preparation of high-quality sequencing libraries is a costly and time-consuming component of metagenomic next generation sequencing (mNGS). While the overall cost of sequencing has dropped significantly over recent years, the reagents needed to prepare sequencing samples are likely to become the dominant expense in the process. Furthermore, libraries prepared by hand are subject to human variability and needless waste due to limitations of manual pipetting volumes. Reduction of reaction volumes, combined with sub-microliter automated dispensing of reagents without consumable pipette tips, has the potential to provide significant advantages. Here, we describe the integration of several instruments, including the Labcyte Echo 525 acoustic liquid handler and the iSeq and NovaSeq Illumina sequencing platforms, to miniaturize and automate mNGS library preparation, significantly reducing the cost and the time required to prepare samples. Through the use of External RNA Controls Consortium (ERCC) spike-in RNAs, we demonstrated the fidelity of the miniaturized preparation to be equivalent to full volume reactions. Furthermore, detection of viral and microbial species from cell culture and patient samples was also maintained in the miniaturized libraries. For 384-well mNGS library preparations, we achieved cost savings of over 80% in materials and reagents alone, and reduced preparation time by 90% compared to manual approaches, without compromising quality or representation within the library.
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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.