ArticleBMC biotechnology2017
Improved genome sequencing using an engineered transposase.
Article in BMC biotechnology, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed, 47 citations in OpenAlex.
- A practical guide to targeted single-cell RNA sequencing technologies.Communications biology · 2026Review
- Strong bias in long-read sequencing prevents assembly ofGenome research · 2026Article
- A novel method for effectively selecting fragments not associated with restriction sites for whole-genome genotyping.BMC biology · 2025Article
- Targeted Next-Generation Sequencing in Rare Diseases.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Emerging trends in genome integration tools for precision engineering of diverse bacterial species.Synthetic biology (Oxford, England) · 2025Review
- OpenTn5: Open-Source Resource for Robust and Scalable Tn5 Transposase Purification and Characterization.bioRxiv : the preprint server for biology · 2024Article
- Low-input PacBio sequencing generates high-quality individual fly genomes and characterizes mutational processes.Nature communications · 2024Article
- Ultrafine mapping of chromosome conformation at hundred basepair resolution reveals regulatory genome architecture.Proceedings of the National Academy of Sciences of the United States of America · 2023Article
- Cost-conscious generation of multiplexed short-read DNA libraries for whole-genome sequencing.PloS one · 2023Article
- Chromatin accessibility profiling by ATAC-seq.Nature protocols · 2022Review
- A single-cell regulatory map of postnatal lung alveologenesis in humans and mice.Cell genomics · 2022Article
- Comprehensive understanding of Tn5 insertion preference improves transcription regulatory element identification.NAR genomics and bioinformatics · 2021Article
- Tagmentation-based single-cell genomics.Genome research · 2021Review
- ReSeq simulates realistic Illumina high-throughput sequencing data.Genome biology · 2021Article
- Angular Leaf Spot ResistanceFrontiers in plant science · 2021Article
- Insertion Specificity of the hATx-6 Transposase ofFrontiers in molecular biosciences · 2021Article
- Development of a sequencing system for spatial decoding of DNA barcode molecules at single-molecule resolution.Communications biology · 2020Article
- Unbiased PCR-free spatio-temporal mapping of the mtDNA mutation spectrum reveals brain region-specific responses to replication instability.BMC biology · 2020Article
- Article
- Comparison of the sequencing bias of currently available library preparation kits for Illumina sequencing of bacterial genomes and metagenomes.DNA research : an international journal for rapid publication of reports on genes and genomes · 2019Article
Corrections and comments
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Authors and funding
8 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundNext-generation sequencing (NGS) has transformed genomic research by reducing turnaround time and cost. However, no major breakthrough has been made in the upstream library preparation methods until the transposase-based Nextera method was invented. Nextera combines DNA fragmentation and barcoding in a single tube reaction and therefore enables a very fast workflow to sequencing-ready DNA libraries within a couple of hours. When compared to the traditional ligation-based methods, transposed-based Nextera has a slight insertion bias.
resultsHere we present the discovery of a mutant transposase (Tn5-059) with a lowered GC insertion bias through protein engineering. We demonstrate Tn5-059 reduces AT dropout and increases uniformity of genome coverage in both bacterial genomes and human genome. We also observe higher library diversity generated by Tn5-059 when compared to Nextera v2 for human exomes, which leads to less sequencing and lower cost per genome. In addition, when used for human exomes, Tn5-059 delivers consistent library insert size over a range of input DNA, allowing up to a tenfold variance from the 50 ng input recommendation.
conclusionsEnhanced DNA input tolerance of Tn5-059 can translate to flexibility and robustness of workflow. DNA input tolerance together with superior uniformity of coverage and lower AT dropouts extend the applications of transposase based library preps. We discuss possible mechanisms of improvements in Tn5-059, and potential advantages of using the new mutant in varieties of applications including microbiome sequencing and chromatin profiling.
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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.