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Automation of PacBio SMRTbell NGS library preparation for bacterial genome sequencing

Overview of attention for article published in Environmental Microbiome, March 2017
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  • Above-average Attention Score compared to outputs of the same age and source (57th percentile)

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Title
Automation of PacBio SMRTbell NGS library preparation for bacterial genome sequencing
Published in
Environmental Microbiome, March 2017
DOI 10.1186/s40793-017-0239-1
Pubmed ID
Authors

Nguyet Kong, Whitney Ng, Kao Thao, Regina Agulto, Allison Weis, Kristi Spittle Kim, Jonas Korlach, Luke Hickey, Lenore Kelly, Stephen Lappin, Bart C. Weimer

Abstract

The PacBio RS II provides for single molecule, real-time DNA technology to sequence genomes and detect DNA modifications. The starting point for high-quality sequence production is high molecular weight genomic DNA. To automate the library preparation process, there must be high-throughput methods in place to assess the genomic DNA, to ensure the size and amounts of the sheared DNA fragments and final library. The library construction automation was accomplished using the Agilent NGS workstation with Bravo accessories for heating, shaking, cooling, and magnetic bead manipulations for template purification. The quality control methods from gDNA input to final library using the Agilent Bioanalyzer System and Agilent TapeStation System were evaluated. Automated protocols of PacBio 10 kb library preparation produced libraries with similar technical performance to those generated manually. The TapeStation System proved to be a reliable method that could be used in a 96-well plate format to QC the DNA equivalent to the standard Bioanalyzer System results. The DNA Integrity Number that is calculated in the TapeStation System software upon analysis of genomic DNA is quite helpful to assure that the starting genomic DNA is not degraded. In this respect, the gDNA assay on the TapeStation System is preferable to the DNA 12000 assay on the Bioanalyzer System, which cannot run genomic DNA, nor can the Bioanalyzer work directly from the 96-well plates.

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Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 42 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
United States 1 2%
Unknown 41 98%

Demographic breakdown

Readers by professional status Count As %
Student > Master 8 19%
Student > Ph. D. Student 7 17%
Researcher 3 7%
Student > Doctoral Student 2 5%
Other 2 5%
Other 4 10%
Unknown 16 38%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 8 19%
Engineering 3 7%
Agricultural and Biological Sciences 3 7%
Immunology and Microbiology 3 7%
Medicine and Dentistry 2 5%
Other 7 17%
Unknown 16 38%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 1. 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 28 March 2017.
All research outputs
#17,289,387
of 25,382,440 outputs
Outputs from Environmental Microbiome
#493
of 786 outputs
Outputs of similar age
#206,789
of 322,668 outputs
Outputs of similar age from Environmental Microbiome
#3
of 7 outputs
Altmetric has tracked 25,382,440 research outputs across all sources so far. This one is in the 21st percentile – i.e., 21% of other outputs scored the same or lower than it.
So far Altmetric has tracked 786 research outputs from this source. They receive a mean Attention Score of 4.8. This one is in the 30th percentile – i.e., 30% of its peers scored the same or lower than it.
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 322,668 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 27th percentile – i.e., 27% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 7 others from the same source and published within six weeks on either side of this one. This one has scored higher than 4 of them.