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 settle for the average answers offered by bulk analysis when you can interrogate toxicity at single-cell resolution?

Juan Garaycoechea

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I am setting up my own lab and BioSkryb has been extremely supportive. We can use PTA to detect mutation with high-confidence, sensitivity, and accuracy in single cells. This really opens up a lot of interesting avenues for us, and places us in the right position to discover some very exciting new biology.”

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Group leader, Hubrecht Institute

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Toxicology, the “science of safety”, is concerned with the detection, nature, effects, treatment, and prevention of harm to living organisms and the environment. Whether the hazard is a chemical substance, biological toxin, or the off-target effects of CRISPR-based gene editing, sensitive detection of subtle intracellular changes and precise identification of perturbed cell populations greatly facilitate the elucidation of toxicity pathways and enable more effective mitigation strategies.

Advances in single-cell isolation techniques, next-generation sequencing, and bioinformatic analysis are enabling precision toxicology, thereby overcoming the limitations of traditional bulk analysis methods. Whilst single-cell RNA-seq is increasingly applied in toxicological studies, routine use of single-cell DNA analysis has been hampered by the lack of reliable whole-genome amplification ([WGA](/products/)) methods.

BioSkryb’s robust single-cell WGA technology ([PTA](/technology/)) and powerful bioinformatics platform ([BaseJumper™](/basejumper/)) enables accurate profiling of mutations induced by chemical or environmental mutagens within and between individual cells. The improved data quality and scalability of PTA also makes it possible to assess both on- and off-target gene editing events across an entire genome at single-cell resolution.