Low-Level Somatic Variant Detection in Tumor FFPE Samples by Sanger Sequencing

Molecular profiling of cancers is becoming more and more important in the research area of personalized cancer therapy, or precision oncology. Next-generation sequencing (NGS) provides a valuable method for high-throughput applications when many targets and samples need to be multiplexed and screened. However, for very focused applications such as single-analyte or single-gene targets, there is a clear need for fast, simple, and affordable methods for detecting low-level somatic variants. The improved sensitivity achieved using Applied Biosystems Minor Variant Finder Software makes Sanger sequencing an ideal technology for filling this gap.

Details of this application of Sanger sequencing can be found in the application note featured below, which demonstrates:

  • The power of high-sensitivity Sanger sequencing in one of its ideal applications: oncology research
  • The detection of low-level (down to 5%) somatic variants using Sanger sequencing along with Minor
    Variant Finder Software
  • The utility of Sanger sequencing for first-line screening in addition to its role as the gold-standard confirmatory
    method for NGS
  • The successful interrogation of minimal amounts (as low as 0.1 ng) of DNA from formalin-fixed,
    paraffin-embedded (FFPE) samples
  • The advantages of Sanger sequencing, including simple workflow and data analysis, low cost per sample (at a limited number of targets), and fast turnaround time

Application Note: Low-level somatic variant detection in tumor FFPE samples by Sanger sequencing

This study demonstrates how the improved sensitivity achieved through Minor Variant Finder Software makes Sanger sequencing an ideal choice for oncology research applications.

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Sanger sequencing workflow

Our simple and fast Sanger sequencing workflow can be completed in less than one workday, from sample to answer. We offer Applied Biosystems products to support many steps of our recommended workflow, from PCR amplification to data analysis. Learn more about each step of our recommended Sanger sequencing workflow.


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