1. Isolation and Quantification of Plasma Circulating Tumor DNA from Melanoma Patients.
- Author
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Marsavela G, Reid A, Gray ES, and Calapre L
- Subjects
- Biomarkers, Tumor blood, Biomarkers, Tumor genetics, DNA, Neoplasm genetics, DNA, Neoplasm isolation & purification, Humans, Melanoma genetics, Melanoma pathology, Plasma metabolism, Proto-Oncogene Proteins B-raf genetics, Circulating Tumor DNA blood, DNA, Neoplasm blood, Melanoma blood, Polymerase Chain Reaction methods
- Abstract
In recent years, circulating tumor DNA (ctDNA) has emerged as a promising prognostic and monitoring biomarker of various cancers, including melanoma. However, sensitive methods are required for its preservation, isolation, and detection. Here we describe a sensitive method for plasma ctDNA isolation using a column-based extraction kit, followed by quantification using a single mutational target with a droplet digital PCR system. This sensitive protocol has been successfully used to quantify diverse mutations present in plasma-derived ctDNA from cancer patients. The full procedure, from blood processing to the analysis of results, takes approximately a day of work.
- Published
- 2021
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