Identification and quantification of heterogeneously-methylated DNA fragments using epiallele-sensitive droplet digital polymerase chain reaction (EAST-ddPCR)
Research output: Contribution to journal › Research article › Contributed › peer-review
Contributors
Abstract
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.
Details
Original language | English |
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Pages (from-to) | 299-312 |
Number of pages | 14 |
Journal | Cancer Genomics and Proteomics |
Volume | 15 |
Issue number | 4 |
Publication status | Published - 1 Jul 2018 |
Peer-reviewed | Yes |
External IDs
PubMed | 29976635 |
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ORCID | /0000-0002-3549-2477/work/148145340 |
Keywords
Sustainable Development Goals
ASJC Scopus subject areas
Keywords
- Digital PCR, DNA methylation, Heterogeneously methylated epialleles, Leukemia, Prostate cancer cells