No-Amp targeted sequencing

Enrichment of hard-to-amplify genomic regions like repeat expansions is now possible with our no-amplification (No-Amp) targeted sequencing method utilizing the CRISPR/Cas9 system.

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Unraveling repeat expansion disorders

Many of the disease-causative genes for repeat expansion disorders were mapped decades ago; however, the underlying disease mechanisms are still not fully understood. These expansions can be several kilobases in size which makes them inaccessible with base-level resolution to most technologies. Now, by combining No-Amp targeted enrichment using the CRISPR/Cas9 system with SMRT Sequencing, scientists can holistically in one experiment:

  • Eliminate PCR bias and errors
  • Sequence through entire repeat expansions with base-level resolution
  • Quantify repeat numbers in normal- and mutant-expanded alleles
  • Identify interruption sequences
  • Characterize somatic mosaicism

Workflow: from gDNA to Complete Repeat Expansion Sequence

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Sample and library prep

With our 2-day No-Amp protocol you can now enrich for your region of interest without amplification using the CRISPR-Cas9 system.

Pacific Biosciences does not sell a kit for carrying out the overall No-Amp Targeted Sequencing method. Use of the method may require rights to third-party owned intellectual property.

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Sequencing

Use the Sequel Systems to accurately sequence a human genome.

  • Sequence multiplexed targets and/or samples on the Sequel Systems using the latest chemistry*
    • Adjust movie time parameters based on repeat expansion size
    • Run up to 48 samples and up to 15 targets per SMRT Cell on the Sequel II or IIe System

*Read lengths, reads/data per SMRT Cell and other sequencing performance results vary based on sample quality/type and insert size.

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Data analysis

Analysis solutions for every user in the lab with SMRT Link and visualization tools.

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Application Brief

Learn more about these best practices for No-Amp targeted sequencing.

Application Brief: Learn more about these best practices for No-Amp targeted sequencing.

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Spotlight

No-Amp targeted sequencing advances Ataxia research

Scientists are exploring the genetic composition of complete repeat expansions to uncover novel phenotype-genotype correlations between Parkinson’s disease and the gene ATXN10. Explore this research further.

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