Detection of Methylated DNA on a Power-Free Microfluidic Chip with Laminar Flow-Assisted Dendritic Amplification

Abstract

We report on a detection method for methylated DNA on a microfluidic chip, which needs no external power for fluid pumping. The methylated DNA was sandwiched by immobilized probe DNA and an anti-methylcytosine antibody. The fluorescence signal was amplified by our original amplification technology. The detection method was first optimized using a 22-mer DNA sequence, then further validated using a 60-mer DNA sequence adapted from the SEPT9 gene. We were able to detect the methylated 60-mer DNA at 0.4 nM within 18 min.

Development of the Bioluminescent Immunoassay for the Detection of 5-Hydroxymethylcytosine in Dinoflagellate

“…This enzyme is relatively stable, and produces a photon by oxidation that is different from the fluorescent detection. 7 So far, we have used this luciferase as a reporter either in a gene expression reporter assay or in an immunoassay accompanied by an antibody. 8,9 The biotinylated Cluc has been shown to be a convenient analytical tool for evaluations of monoclonal antibody localization in a living body.…”

Fast and Single-step Fluorescence-based Competitive Bioassay Microdevice Combined PDMS Microchannel Arrays Separately Immobilizing Graphene Oxide-Analyte Conjugates and Fluorescently-labelled Receptor Proteins

“…However, long analysis time, high sample volume, and complicated operation are still recognized as the problems to be solved by many researchers in this field. [1][2][3][4] Recently, a large number of studies on technological developments of microsystems, microchemistry, and fully automated analytical systems have been reported in attempts to solve these problems. [5][6][7][8] However, simultaneously solving these problems is still challenging.…”

Effect of DNA Methylation on the Velocity of DNA Translocation through a Nanochannel

“…1 It is well known that DNA methylation is an important regulator of epigenetic alterations, and increased methylation has been detected in patients with cancers. 2,3 Therefore, a better understanding of DNA methylation is desirable for improving the diagnosis and prognosis of cancer and prediction of responses to many kinds of cancer therapies. 4 Although understanding how DNA methylation affects DNA flexibility is important for understanding the role of DNA methylation in regulating chromosome packaging and gene expression at the molecular scale, the effect of DNA methylation on the physical properties of DNA remains poorly understood.…”