Issue 3, 2019

Multilayer microfluidic array for highly efficient sample loading and digital melt analysis of DNA methylation

Abstract

Liquid biopsies contain a treasure of genetic and epigenetic biomarkers that contain information for the detection and monitoring of human disease. DNA methylation is an epigenetic modification that is critical to determining cellular phenotype and often becomes altered in many disease states. In cancer, aberrant DNA methylation contributes to carcinogenesis and can profoundly affect tumor evolution, metastatic potential, and resistance to therapeutic intervention. However, current technologies are not well-suited for quantitative assessment of DNA methylation heterogeneity, especially in challenging samples such as liquid biopsies with low DNA input and high background. We present a multilayer microfluidic device for quantitative analysis of DNA methylation by digital PCR and high resolution melt (HRM). The multilayer design facilitates high-density array digitization aimed at maximizing sample loading efficiency. The platform achieves highly parallelized digital PCR-HRM-based discrimination of rare heterogeneous DNA methylation as low as 0.0001% methylated/unmethylated molecules of a classic tumor suppressor gene, CDKN2A (p14ARF).

Graphical abstract: Multilayer microfluidic array for highly efficient sample loading and digital melt analysis of DNA methylation

Supplementary files

Article information

Article type
Paper
Submitted
31 اکتوٗبر 2018
Accepted
28 دسمبر 2018
First published
09 جنؤری 2019

Lab Chip, 2019,19, 444-451

Multilayer microfluidic array for highly efficient sample loading and digital melt analysis of DNA methylation

C. M. O'Keefe, D. Giammanco, S. Li, T. R. Pisanic and T. J. Wang, Lab Chip, 2019, 19, 444 DOI: 10.1039/C8LC01189C

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements