Primary Template-directed Amplification or PTA employs controlled reaction parameters to reproducibly recover greater than 95% of the genome of single-cells and severely limited input samples with best-in-class fidelity and uniformity. 

Discover the difference BioSkryb Genomics can make for you in WGA

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Improvements in whole genome amplification (WGA) would enable new types of basic and applied biomedical research, including studies of intratissue genetic diversity that require more accurate single-cell genotyping. Here, we present primary template-directed amplification (PTA), an isothermal WGA method that reproducibly captures >95% of the genomes of single cells in a more uniform and accurate manner than existing approaches, resulting in significantly improved variant calling sensitivity and precision. To illustrate the types of studies that are enabled by PTA, we developed direct measurement of environmental mutagenicity (DMEM), a tool for mapping genome-wide interactions of mutagens with single living human cells at base-pair resolution. In addition, we utilized PTA for genome-wide off-target indel and structural variant detection in cells that had undergone CRISPR-mediated genome editing, establishing the feasibility for performing single-cell evaluations of biopsies from edited tissues. The improved precision and accuracy of variant detection with PTA overcomes the current limitations of accurate WGA, which is the major obstacle to studying genetic diversity and evolution at cellular resolution.

The improved precision and accuracy of variant detection with PTA overcomes the current limitations of accurate WGA, which is the major obstacle to studying genetic diversity and evolution at cellular resolution.

Accurate Genomic Variant Detection in Single Cells with Primary Template-Directed Amplification

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## Why rely on culturing and complex metagenomic analyses if you can resolve a bacterial genome from a single cell?

Bacteria make up the bulk of life on earth, and it is estimated that humans are aware of only a small percentage of microbes. Even with our limited knowledge of bacterial diversity around us, ground breaking scientific discoveries have been made possible because of the bacteria around us, including Taq polymerase, restriction endonucleases, and CRISPR gene editing technology. Additionally, the more we learn about the symbiotic relationship between humans and bacteria, the more we appreciate the profound impact bacteria have on human health.

ResolveDNA Microbiome unlocks the near-complete genome of a single bacterium, enabling myriad new applications.

Genomic Analysis of Single Bacteria using ResolveDNA Microbiome

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Genomic Analysis of Single Bacteria using ResolveDNA® Microbiome

The causes and consequences of microbial variation across biomes have been the subject of intense study for over a century. Specifically, bacterial niche partitioning has been shown to play fundamental roles in the structuring and maintenance of global biogeochemical cycles. Additionally, eukaryotic organisms are inhabited by complex microbial communities, collectively named the microbiota, that have been shown to modulate diverse host physiological traits ultimately impacting the host fitness. A bottleneck to unraveling the vast uncharted bacterial diversity across biomes has been the lack of robust technology to faithfully amplify the femtogram quantities of DNA of a single bacterium. Here we present ResolveDNA Microbiome, a whole-genome amplification technology, to reconstruct high-quality genomes of single bacteria

Here we present ResolveDNA Microbiome, a whole-genome amplification technology, to reconstruct high-quality genomes of single bacteria.

Charting and deconvolution of microbial diversity by high-quality single-cell genomic reconstruction utilizing ResolveDNA Microbiome

ASM Microbe 2022

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Charting and deconvolution of microbial diversity by high-quality single-cell genomic reconstruction utilizing ResolveDNA Microbiome 

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Microorganisms are ubiquitous, representing the most abundant—but least characterized—group of life on Earth. Their innumerable and intricate symbiotic and pathogenic relationships have a profound impact on human well-being and global sustainability.

Microbes have a critical impact on the health of their hosts in the context of diseases caused by single infectious agents, or the complex biology of our gut microbiomes. In addition, they have provided us with therapeutic biomolecules and tools for food production, bioprocessing and molecular biology. Nevertheless, their genetic material remains a largely untapped resource. The discovery and characterization of microbial genomes continues to be hampered by an inability to recover sufficient material for bulk whole genome sequencing.

BioSkryb’s versatile and scalable single-cell whole genome amplification ([WGA](/products/)) technology ([PTA](/technology/)) and powerful bioinformatics platform ([BaseJumper™](/basejumper/)) removes a critical bottleneck in microbiome research and bioprospecting by enabling the recovery of high-quality, near-complete genomes from individual bacterial cells, whether cultured or from environmental/clinical metagenomic samples. As such, PTA promises to advance discovery of novel microbial species, and unlock our understanding of microbial diversity and functional genomics for a wide range of applications in microbiome research, medicine, agriculture, and biotechnology.

Microbiome Research

Why rely on culturing and complex metagenomic analyses if you can resolve a bacterial genome from a single cell?

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