NSTI BioNano 2010

Integrated polymer-based microfluidic device for DNA preparation in pyrosquencer

P. Khuntontong, C.H. Kua, M. Gong, I.S.H. Foo, S. Goel, Z.P. Wang
Singapore Institute of Manufacturing Technology, SG

Keywords: DNA preparation, silica beads purification, lap on a chip, polymer microfluidic

Abstract:

We present a polymer-based microfluidic device capable of performing various DNA preparation processes like enzymatic, end repairing, and purification. The system consisted of a disposable polymeric chip and a controlling unit. Mixing chambers, incubation chambers and DNA purification modules were integrated on the chip. The channels on the polymeric chip were designed to be in spiral-shape to increase mixing efficiency. The channels were treated with polyurethane to create a hydrophobic surface, which helped to minimize the residual liquid. The regulation of the temperature and the control of the fluidic flow on the polymeric chip were performed by the controlling unit. The entire DNA processing needed 80 minutes. Genome from HCT116 cells and pUC19 DNA were used. The end-repairing process and digestion by enzyme were performed on DNA at 37°C for an hour in the channels. DNA sample were constantly shuffle around the first and the second spiral-shape chamber to ensure a homogeneous mixing of DNA and digestive enzyme. After the enzymatic process, silica purification technique was used to purify the DNA. We have separately optimized the purification process by studying the effect of mixing time and velocity, temperature, pH, and concentration of solution. The experiment results and suggestions are reported.
 
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