A sample-to-answer droplet magnetofluidic assay platform for quantitative methylation-specific PCR

  • Alejandro Stark
  • Dong Jin Shin
  • Tza-Huei WangEmail author


Dysregulation of DNA methylation has been identified as an epigenetic biomarker for numerous cancer types. Gene-specific identification techniques relying on methylation-specific PCR (MSP) require a lengthy manual benchtop process that is susceptible to human-error and contamination. This MSP assay requires a series of discrete sample processing steps including genomic DNA extraction, bisulfite conversion and readout via PCR. In this work, we present a streamlined assay platform utilizing droplet magnetofluidic principles for integration of all sample processing steps required to obtain quantitative MSP signal from raw biological samples. We present a streamlined protocol for solid-phase extraction and bisulfite conversion of genomic DNA, which minimizes reagent use and simplifies the sample preparation protocol for implementation on a compact assay platform. Furthermore, we present a thermally robust assay chip that enables DNA extraction, bisulfite conversion and quantitative PCR from biological samples on a single device. Technical improvements to facilitate DNA extraction and PCR on a single chip in addition to chip performance characterization data are presented.


Methylation Epigenetics Methylation-specific PCR Lab-on-chip Magnetofluidics qPCR Biomarkers 



The authors would also like to thank funding sources from National Institutes of Health (R21CA186809, UG3CA211457, U01CA200499).


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© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Alejandro Stark
    • 1
  • Dong Jin Shin
    • 1
  • Tza-Huei Wang
    • 1
    • 2
    • 3
    Email author
  1. 1.Department of Mechanical EngineeringJohns Hopkins UniversityBaltimoreUSA
  2. 2.Department of Biomedical EngineeringJohns Hopkins UniversityBaltimoreUSA
  3. 3.Johns Hopkins Institute for NanoBioTechnologyBaltimoreUSA

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