BioTechniques (Aug 2018)

Suitability of melanoma FFPE samples for NGS libraries: time and quality thresholds for downstream molecular tests

  • David Millán-Esteban,
  • Diana Reyes-García,
  • Zaida García-Casado,
  • José Bañuls,
  • José Antonio López-Guerrero,
  • Celia Requena,
  • Aranzazu Rodríguez-Hernández,
  • Víctor Traves,
  • Eduardo Nagore

DOI
https://doi.org/10.2144/btn-2018-0016
Journal volume & issue
Vol. 65, no. 2
pp. 79 – 85

Abstract

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The use of NGS in clinical practice for precision diagnosis requires a quality starting material. Despite the broadly established use of formalin-fixed paraffin-embedded (FFPE) samples in molecular testing, these usually have low-quality DNA. We established a method to determine the suitability of melanoma FFPE samples for an amplicon-based NGS custom panel analysis. DNA was extracted from unstained melanoma samples and wide local excision samples. Amplicon-based libraries were constructed and tested using time and quality parameters as variables. Time elapsed from sample retrieval >7 years, a quality control value > 5.63 and a DNA integrity value < 2.05 indicated samples were not suitable. A decision tree is provided with rate of samples suitable for analysis according to the combination of these parameters.

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