ArticlePloS one2016
Digital Droplet Multiple Displacement Amplification (ddMDA) for Whole Genome Sequencing of Limited DNA Samples.
Article in PloS one, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Who cites it
27 citing papers in PubMed.
- Advances in single-cell long-read sequencing technologies.NAR genomics and bioinformatics · 2024Article
- Droplet based whole genome amplification for sequencing minute amounts of purified Mycobacterium tuberculosis DNA.Scientific reports · 2024Article
- Impact of template denaturation prior to whole genome amplification on gene detection in high GC-content species, Burkholderia mallei and B. pseudomallei.BMC research notes · 2024Article
- Evaluation of multiple displacement amplification for metagenomic analysis of low biomass samples.ISME communications · 2024Article
- Back to Basics: A Simplified Improvement to Multiple Displacement Amplification for Microbial Single-Cell Genomics.International journal of molecular sciences · 2023Article
- Ultralow-Input Genome Library Preparation for Nanopore Sequencing with Droplet MDA.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Cell-Free Production Systems in Droplet Microfluidics.Advances in biochemical engineering/biotechnology · 2023Review
- COMicromachines · 2022Article
- Isolation and sequencing of a single copy of an introgressed chromosome from a complex genome for gene and SNP identification.TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik · 2022Article
- Surveillance of Listeria monocytogenes: Early Detection, Population Dynamics, and Quasimetagenomic Sequencing during Selective Enrichment.Applied and environmental microbiology · 2021Article
- Quantifying genome DNA during whole-genome amplificationRSC advances · 2021Article
- Comparison of Multiple Displacement Amplification (MDA) and Multiple Annealing and Looping-Based Amplification Cycles (MALBAC) in Limited DNA Sequencing Based on Tube and Droplet.Micromachines · 2020Article
- Obtaining Genome Sequences of Mutualistic Bacteria in SingleInternational journal of molecular sciences · 2019Article
- Bacterial Single Cell Whole Transcriptome Amplification in Microfluidic Platform Shows Putative Gene Expression Heterogeneity.Analytical chemistry · 2019Article
- Recent Advances in Droplet-based Microfluidic Technologies for Biochemistry and Molecular Biology.Micromachines · 2019Review
- Droplet microfluidics for high-sensitivity and high-throughput detection and screening of disease biomarkers.Wiley interdisciplinary reviews. Nanomedicine and nanobiotechnology · 2018Review
- The Development of an Effective Bacterial Single-Cell Lysis Method Suitable for Whole Genome Amplification in Microfluidic Platforms.Micromachines · 2018Article
- Evaluation of bias induced by viral enrichment and random amplification protocols in metagenomic surveys of saliva DNA viruses.Microbiome · 2018Article
- PaSD-qc: quality control for single cell whole-genome sequencing data using power spectral density estimation.Nucleic acids research · 2018Article
- Obtaining high-quality draft genomes from uncultured microbes by cleaning and co-assembly of single-cell amplified genomes.Scientific reports · 2018Article
Corrections and comments
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Authors and funding
4 authors.
Funding
Abstract
Multiple displacement amplification (MDA) is a widely used technique for amplification of DNA from samples containing limited amounts of DNA (e.g., uncultivable microbes or clinical samples) before whole genome sequencing. Despite its advantages of high yield and fidelity, it suffers from high amplification bias and non-specific amplification when amplifying sub-nanogram of template DNA. Here, we present a microfluidic digital droplet MDA (ddMDA) technique where partitioning of the template DNA into thousands of sub-nanoliter droplets, each containing a small number of DNA fragments, greatly reduces the competition among DNA fragments for primers and polymerase thereby greatly reducing amplification bias. Consequently, the ddMDA approach enabled a more uniform coverage of amplification over the entire length of the genome, with significantly lower bias and non-specific amplification than conventional MDA. For a sample containing 0.1 pg/μL of E. coli DNA (equivalent of ~3/1000 of an E. coli genome per droplet), ddMDA achieves a 65-fold increase in coverage in de novo assembly, and more than 20-fold increase in specificity (percentage of reads mapping to E. coli) compared to the conventional tube MDA. ddMDA offers a powerful method useful for many applications including medical diagnostics, forensics, and environmental microbiology.
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Registered trials
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