Evidence map›Paper›PMID 29859085›Full record

ArticleBMC biotechnology2018

DNA assembly with error correction on a droplet digital microfluidics platform.

Yuliya Khilko, Philip D Weyman, John I Glass, Mark D Adams, Melanie A McNeil, Peter B Griffin

Abstract read
In one paragraph

Article in BMC biotechnology, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.

0numbers the graph read from it
0cells of the map it votes in
4citing papers in PubMed
–field-weighted citation impact
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

4 citing papers in PubMed.

  1. Article
  2. DNA storage: The future direction for medical cold data storage.Synthetic and systems biotechnology · 2025
    Review
  3. DNA-DISK: Automated end-to-end data storage via enzymatic single-nucleotide DNA synthesis and sequencing on digital microfluidics.Proceedings of the National Academy of Sciences of the United States of America · 2024
    Article
  4. Review
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

6 authors.

Yuliya KhilkoStanford Genome Technology Center, Stanford University, 3165 Porter Drive, Palo Alto, CA, 94304, USA.
Philip D WeymanJ. Craig Venter Institute, 4120 Capricorn Lane, La Jolla, CA, 92037, USA.
John I GlassJ. Craig Venter Institute, 4120 Capricorn Lane, La Jolla, CA, 92037, USA.
Mark D AdamsJ. Craig Venter Institute, 4120 Capricorn Lane, La Jolla, CA, 92037, USA.
Melanie A McNeilDepartment of Biomedical, Chemical and Materials Engineering, San Jose State University, 1 Washington Sq, San Jose, CA, 95192, USA.
Peter B GriffinStanford Genome Technology Center, Stanford University, 3165 Porter Drive, Palo Alto, CA, 94304, USA. griffin@stanford.edu.

Funding

A Digital Microfluidic Systems for Gene Synthesis, Sequencing and RecoveryR21GM104694 · NIGMS · J. CRAIG VENTER INSTITUTE, INC. · PI GLASS, JOHN IRVIN · 2012 to 2014
$697k
NIGMS NIH HHS 5R21GM104694-03NIGMS NIH HHS R21 GM104694
6 · The paper itself

Abstract

backgroundCustom synthesized DNA is in high demand for synthetic biology applications. However, current technologies to produce these sequences using assembly from DNA oligonucleotides are costly and labor-intensive. The automation and reduced sample volumes afforded by microfluidic technologies could significantly decrease materials and labor costs associated with DNA synthesis. The purpose of this study was to develop a gene assembly protocol utilizing a digital microfluidic device. Toward this goal, we adapted bench-scale oligonucleotide assembly methods followed by enzymatic error correction to the Mondrian™ digital microfluidic platform.

resultsWe optimized Gibson assembly, polymerase chain reaction (PCR), and enzymatic error correction reactions in a single protocol to assemble 12 oligonucleotides into a 339-bp double- stranded DNA sequence encoding part of the human influenza virus hemagglutinin (HA) gene. The reactions were scaled down to 0.6-1.2 μL. Initial microfluidic assembly methods were successful and had an error frequency of approximately 4 errors/kb with errors originating from the original oligonucleotide synthesis. Relative to conventional benchtop procedures, PCR optimization required additional amounts of MgCl

conclusionWe demonstrated that DNA assembly from oligonucleotides and error correction could be completely automated on a digital microfluidic (DMF) platform. The results demonstrate that enzymatic reactions in droplets show a strong dependence on surface interactions, and successful on-chip implementation required supplementation with surfactants, molecular crowding agents, and an excess of enzyme. Enzymatic error correction of assembled fragments improved sequence fidelity by 2-fold, which was a significant improvement but somewhat lower than expected compared to bench-top assays, suggesting an additional capacity for optimization.

Indexed as

DNA, ViralHemagglutinin Glycoproteins, Influenza VirusHumansInfluenza A Virus, H9N2 SubtypeInfluenza, HumanMicrofluidic Analytical TechniquesMicrofluidicsOligonucleotide Array Sequence AnalysisPolymerase Chain ReactionDNA, ViralHemagglutinin Glycoproteins, Influenza Virushemagglutinin HA1, influenza B virusDigital microfluidicsDNAError correctionGibson assembly

Identifiers

PMID29859085
PMCPMC5984785

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Registered trials

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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.