Evidence map›Paper›PMID 28095828›Full record

ArticleBMC biotechnology2017

Improved genome sequencing using an engineered transposase.

Amirali Kia, Christian Gloeckner, Trina Osothprarop, Niall Gormley, Erin Bomati, Michelle Stephenson, Igor Goryshin, Molly Min He

Open access · goldAbstract read
In one paragraph

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.

0numbers the graph read from it
0cells of the map it votes in
23citing papers in PubMed
9.6field-weighted citation impact, top 3% of its field
1 · What the graph read from it

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.

2 · The registry

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.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

23 citing papers in PubMed, 47 citations in OpenAlex.

  1. Review
  2. Article
  3. Article
  4. Targeted Next-Generation Sequencing in Rare Diseases.Methods in molecular biology (Clifton, N.J.) · 2025
    Article
  5. Review
  6. Article
  7. Article
  8. 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 · 2023
    Article
  9. Article
  10. Review
  11. Article
  12. Article
  13. Review
  14. Article
  15. Angular Leaf Spot ResistanceFrontiers in plant science · 2021
    Article
  16. Insertion Specificity of the hATx-6 Transposase ofFrontiers in molecular biosciences · 2021
    Article
  17. Article
  18. Article
  19. Article
  20. 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 · 2019
    Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

8 authors at 2 institutions in 2 countries.

Amirali KiaDepartment of Protein Engineering, Illumina Inc, 5200 Illumina Way, San Diego, CA, USA.
Christian GloecknerNEO New Oncology GmbH, Gottfried-Hagen-Str. 20, Cologne, 51105, Germany.
Trina OsothpraropDepartment of Protein Engineering, Illumina Inc, 5200 Illumina Way, San Diego, CA, USA.
Niall GormleyTechnology Development, Illumina Inc, Little Chesterford, Nr Saffron Walden, Essex, CB10 1XL, UK.
Erin BomatiDepartment of Protein Engineering, Illumina Inc, 5200 Illumina Way, San Diego, CA, USA.
Michelle StephensonDepartment of Protein Engineering, Illumina Inc, 5200 Illumina Way, San Diego, CA, USA.
Igor GoryshinIllumina Inc, 5602 Research Park Blvd., Suite 200, Madison, WI, USA.
Molly Min HeDepartment of Protein Engineering, Illumina Inc, 5200 Illumina Way, San Diego, CA, USA. mhe@illumina.com.
Illumina (United States) · USIllumina (United Kingdom) · GB

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

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.

Indexed as

Protein EngineeringAT Rich SequenceChromosome MappingDNAHigh-Throughput Nucleotide SequencingReproducibility of ResultsSensitivity and SpecificitySequence Analysis, DNATransposasesDNATransposases

Identifiers

PMID28095828
PMCPMC5240201
OpenAlexW2572666481

What OpenQuestion holds

Textmetadata
LicenceCC BY
Read underepoch 390

Registered trials

None linked

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.