Evidence map›Paper›PMID 38709890›Full record

ArticleNucleic acids research2024

Arrayed in vivo barcoding for multiplexed sequence verification of plasmid DNA and demultiplexing of pooled libraries.

Weiyi Li, Darach Miller, Xianan Liu, Lorenzo Tosi, Lamia Chkaiban, Han Mei, Po-Hsiang Hung, Biju Parekkadan, Gavin Sherlock, Sasha F Levy

Abstract read
In one paragraph

Article in Nucleic acids research, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.

0numbers the graph read from it
0cells of the map it votes in
7citing 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

7 citing papers in PubMed.

  1. Article
  2. Article
  3. Scaling DNA engineering.Trends in biotechnology · 2025
    Review
  4. Article
  5. Review
  6. Review
  7. Article
4 · The record

Corrections and comments

5 · Who and what money

Authors and funding

10 authors.

Weiyi LiSLAC National Accelerator Laboratory, Stanford University, Stanford, CA, USA.ORCID 0000-0002-1168-7093
Darach MillerSLAC National Accelerator Laboratory, Stanford University, Stanford, CA, USA.
Xianan LiuSLAC National Accelerator Laboratory, Stanford University, Stanford, CA, USA.
Lorenzo TosiDepartment of Biomedical Engineering, Rutgers University, Piscataway, NJ, USA.
Lamia ChkaibanDepartment of Biomedical Engineering, Rutgers University, Piscataway, NJ, USA.
Han MeiSLAC National Accelerator Laboratory, Stanford University, Stanford, CA, USA.
Po-Hsiang HungDepartment of Genetics, Stanford University School of Medicine, Stanford, CA, USA.
Biju ParekkadanDepartment of Biomedical Engineering, Rutgers University, Piscataway, NJ, USA.
Gavin SherlockDepartment of Genetics, Stanford University School of Medicine, Stanford, CA, USA.ORCID 0000-0002-1692-4983
Sasha F LevySLAC National Accelerator Laboratory, Stanford University, Stanford, CA, USA.

Funding

Multiplexed In Vivo DNA AssemblyR01HG011676 · NHGRI · STANFORD UNIVERSITY · PI LEVY, SASHA F · 2022 to 2024
$2.9M
PPiSeq: High-Throughput Protein-Protein Interaction SequencingR01AI164530 · NIAID · STANFORD UNIVERSITY · PI SHERLOCK, GAVIN J · 2021 to 2024
$1.7M
NHGRI NIH HHS R01 HG011676NIAID NIH HHS R01 AI164530NIH HHS R01HG011676
6 · The paper itself

Abstract

Sequence verification of plasmid DNA is critical for many cloning and molecular biology workflows. To leverage high-throughput sequencing, several methods have been developed that add a unique DNA barcode to individual samples prior to pooling and sequencing. However, these methods require an individual plasmid extraction and/or in vitro barcoding reaction for each sample processed, limiting throughput and adding cost. Here, we develop an arrayed in vivo plasmid barcoding platform that enables pooled plasmid extraction and library preparation for Oxford Nanopore sequencing. This method has a high accuracy and recovery rate, and greatly increases throughput and reduces cost relative to other plasmid barcoding methods or Sanger sequencing. We use in vivo barcoding to sequence verify >45 000 plasmids and show that the method can be used to transform error-containing dispersed plasmid pools into sequence-perfect arrays or well-balanced pools. In vivo barcoding does not require any specialized equipment beyond a low-overhead Oxford Nanopore sequencer, enabling most labs to flexibly process hundreds to thousands of plasmids in parallel.

Indexed as

Gene LibraryHigh-Throughput Nucleotide SequencingPlasmidsDNADNA Barcoding, TaxonomicNanopore SequencingSequence Analysis, DNADNA

Identifiers

PMID38709890
PMCPMC11162764

What OpenQuestion holds

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LicenceCC BY
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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.