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Solid phase nucleic acid extraction technique in a microfluidic chip using a novel non-chaotropic agent: dimethyl adipimidate
Citation
Shin, Y and Perera, AP and Wong, CC and Park, MK, Solid phase nucleic acid extraction technique in a microfluidic chip using a novel non-chaotropic agent: dimethyl adipimidate, Lab on A Chip, 14 pp. 359-368. ISSN 1473-0197 (2014) [Refereed Article]
Copyright Statement
Copyright 2014 The Royal Society of Chemistry
Abstract
Here, we present a silicon microfluidic system for the purification and extraction of nucleic acids from human body fluid samples utilizing a dimethyl adipimidate (DMA)-based solid-phase extraction method. We propose DMA, which has been used as an amino-reactive cross-linking agent within cells and proteins, as a non-chaotropic reagent for the capture of nucleic acids to overcome the limitations of existing chaotropic and non-chaotropic techniques such as low binding efficiency, PCR inhibition and so on. DMA contains bi-functional imidoesters that form reversible cross-linking structures with DNA therefore providing a high surface-area to volume ratio for capturing DNA without structurally modifying microfluidic channels. In this work, we have first demonstrated highly efficient capture and purification of genomic DNA (T24 cell line) with DMA using a label-free silicon microring resonator sensor device. In addition, we observed the improvement of the DNA amplification efficiency by using the proposed technique for both the genetic (HRAS) and epigenetic (RARβ) analysis of DNA biomarkers. Particularly, we confirmed that the DMA-based solid-phase extraction technique can be applied for the extraction of genomic DNA with higher purity (p < 0.001) using human body fluids (blood and urine) in silicon microfluidic devices compared to other chaotropic methods. Therefore, the proposed technique would be able to harmonize with a micro-total analysis system platform for the analysis of genetic and epigenetic DNA biomarkers related to human diseases in the field of point-of-care (POC) diagnostic applications.
Item Details
| Item Type: | Refereed Article |
|---|---|
| Keywords: | dimethyl adipimidate, microfluidic chip |
| Research Division: | Biomedical and Clinical Sciences |
| Research Group: | Clinical sciences |
| Research Field: | Medical genetics (excl. cancer genetics) |
| Objective Division: | Health |
| Objective Group: | Clinical health |
| Objective Field: | Clinical health not elsewhere classified |
| UTAS Author: | Perera, AP (Mrs Agampodi Perera) |
| ID Code: | 120566 |
| Year Published: | 2014 |
| Web of Science® Times Cited: | 24 |
| Deposited By: | Ecology and Biodiversity |
| Deposited On: | 2017-08-29 |
| Last Modified: | 2017-10-23 |
| Downloads: | 0 |
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