OCT 01, 2015 12:00 PM PDT

Detection and sequencing of rare mutations using mutation enrichment via COLD-PCR

Speakers
  • Professor and Director, Division of Medical Physics and Biophysics, Director, DNA technology and SARRP pre-clinical laboratories, Brigham and Women's Hospital, Dana-Farber Cancer Institute
    Biography
      Dr. Makrigiorgos is a Professor of Radiation Oncology and Director of the Medical Physics & Biophysics division at Dana Farber Cancer Institute and Brigham and Women's Hospitals, Harvard Medical School. He also directs the DNA technology laboratory and the radiation pre-clinical facility. His research interests include the development of novel DNA technologies for molecular diagnostics in Oncology and he identification of circulating cancer biomarkers. He is the inventor of several PCR-based techniques for molecular diagnostics, including Balanced-PCR, Anti-primer-quenching-real-time PCR, Hairpin-PCR, COLD-PCR and DiSSeCT technology. He is a Member of the Editorial Board of Clinical Chemistry and has published over 120 articles, reviews and book chapters.

    Abstract:

    Detection of rare mutations is required for reliable identification of mutations in liquid biopsies and heterogeneous tumors. Yet low-level mutation detection remains a technically challenging task, especially when the position and type of mutation are unknown. We describe simple methods for enrichment of mutations in a sample, such that downstream detection or sequencing of mutations is facilitated. Using COLD-PCR, the sensitivity of almost all mutation detection methods is improved including targeted sequencing, digital PCR, and high resolution melting. Learning objectives include the adaptation of COLD-PCR to boost the mutation detection sensitivity for all these established platforms; and practical tips to get the most out of the combined technologies.


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