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Improved genome sequencing using an engineered transposase

Overview of attention for article published in BMC Biotechnology, January 2017
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  • In the top 25% of all research outputs scored by Altmetric
  • High Attention Score compared to outputs of the same age (81st percentile)
  • High Attention Score compared to outputs of the same age and source (92nd percentile)

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3 X users
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6 patents

Citations

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31 Dimensions

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156 Mendeley
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Title
Improved genome sequencing using an engineered transposase
Published in
BMC Biotechnology, January 2017
DOI 10.1186/s12896-016-0326-1
Pubmed ID
Authors

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

Abstract

Next-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. Here 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. Enhanced 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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Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 156 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
China 1 <1%
Switzerland 1 <1%
South Africa 1 <1%
Unknown 153 98%

Demographic breakdown

Readers by professional status Count As %
Researcher 34 22%
Student > Ph. D. Student 30 19%
Student > Bachelor 21 13%
Student > Master 13 8%
Student > Postgraduate 7 4%
Other 15 10%
Unknown 36 23%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 46 29%
Agricultural and Biological Sciences 35 22%
Engineering 8 5%
Medicine and Dentistry 7 4%
Neuroscience 5 3%
Other 16 10%
Unknown 39 25%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 8. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 27 October 2022.
All research outputs
#4,085,262
of 24,093,053 outputs
Outputs from BMC Biotechnology
#200
of 951 outputs
Outputs of similar age
#78,837
of 425,086 outputs
Outputs of similar age from BMC Biotechnology
#2
of 14 outputs
Altmetric has tracked 24,093,053 research outputs across all sources so far. Compared to these this one has done well and is in the 82nd percentile: it's in the top 25% of all research outputs ever tracked by Altmetric.
So far Altmetric has tracked 951 research outputs from this source. They typically receive more attention than average, with a mean Attention Score of 7.9. This one has done well, scoring higher than 78% of its peers.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 425,086 tracked outputs that were published within six weeks on either side of this one in any source. This one has done well, scoring higher than 81% of its contemporaries.
We're also able to compare this research output to 14 others from the same source and published within six weeks on either side of this one. This one has done particularly well, scoring higher than 92% of its contemporaries.