TY - JOUR T1 - Identification and Quantification of Heterogeneously-methylated DNA Fragments Using Epiallele-sensitive Droplet Digital Polymerase Chain Reaction (EAST-ddPCR) JF - Cancer Genomics - Proteomics JO - Cancer Genomics Proteomics SP - 299 LP - 312 DO - 10.21873/cgp.20088 VL - 15 IS - 4 AU - MARIO MENSCHIKOWSKI AU - CARSTEN JANDECK AU - MARKUS FRIEDEMANN AU - SUSAN RICHTER AU - DANA THIEM AU - BJÖRN SÖNKE LANGE AU - MEINOLF SUTTORP Y1 - 2018/07/01 UR - http://cgp.iiarjournals.org/content/15/4/299.abstract N2 - Background/Aim: DNA methylation plays an important role in the initiation and propagation of carcinogenesis; however, the role of heterogeneously methylated epialleles is currently not well studied, also due to the lack of sensitive, unbiased and high throughput methods. Here, a newly developed droplet digital PCR (ddPCR)-based method was evaluated regarding its ability to quantify such heterogeneously methylated epialleles with sufficient analytical sensitivity and specificity. Materials and Methods: Genomic DNA from blood leukocytes and bone marrow aspirate of an 8-year old male with B-cell acute lymphoblastic leukemia (B-ALL) and from normal and malignant prostate cell lines were analysed using ddPCR. Results: By using these DNA samples, the specificity of an applied set of fluorescence-labeled probes was demonstrated as a proof of concept. Conclusion: All individual heterogeneously-methylated epialleles were quantifiable by a set of fluorescence-labeled probes with complementary sequences to epialleles in a closed-tube and high-throughput manner. The new method named epiallele-sensitive droplet digital PCR (EAST-ddPCR) may give new insights in the generation and regulation of epialleles and may help in finding new biomarkers for the diagnosis of benign und malignant diseases. ER -