The identification of oncogenic driver mutations has led to the rapid rise of genotype-directed treatments. However, genetic analysis of tumors remains cumbersome and a morbid experience for patients. Noninvasive assessment of tumor genotype, so-called “liquid biopsy,” such as plasma genotyping represents a potentially transformative tool. Here we describe a genotyping protocol of cell-free plasma DNA (cfDNA) using Droplet Digital™ PCR (ddPCR™). ddPCR emulsifies DNA into ~20,000 droplets in which PCR is performed to endpoint in each droplet for both mutant and wild-type DNA. Droplets are run through a modified flow cytometer where mutant and wild-type DNA emit different colored signals. The count of these signals upon Poisson distribution analysis allows sensitive quantification of allelic prevalence.
CITATION STYLE
Kuang, Y., O’Connell, A., Sacher, A. G., Feeney, N., Alden, R. S., Oxnard, G. R., & Paweletz, C. P. (2018). Monitoring of response and resistance in plasma of EGFR-mutant lung cancer using droplet digital PCR. In Methods in Molecular Biology (Vol. 1768, pp. 193–207). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7778-9_12
Mendeley helps you to discover research relevant for your work.