PCR Microfluidic Device for Detection of Low Copy Number Nucleic Acids.
Pierre-Alain Auroux, Felix Niggli, Andreas Manz, Philip Day
Keywords: micro-fluidics, DNA, PCR, lab-on-a-chip
The polymerase chain reaction (PCR) is an enzyme-mediated nucleic acid amplifer process that is employed extensively throughout the medical and biological sciences. The technique has risen to acclaim largely through its exquisite sensitivity, but an inability to distinguish trace contaminants plus high operating costs greatly reduce its impact on further potential applications. Additionally, the process tends to diminish gene detection sensitivity within a highly heterogenous cell population; detection may be totally lost if even marginally diluted. A microfluidics device has intrinsic properties, particularly if multi-parallelised, can alleviate risk of contamination and also increases detection sensitivity of low copy number genes in complex cell populations, which we demonstrate utilizing a PCR amplifier device with shunting microfluidics.
Our reactor consists of a microstructure constructed from SU-8, and sandwiched by PMMA. The proof of principle experiment was conducted with a modified glass capillary manufactured by Roche Diagnostics GmbH. Firstly PCR was performed without any optimization and the sample was then analyzed using an Agilent 2100 Bioanalyser. The specifically amplified product was determined to have a concentration of 0.36ng/uL. The analysis of positive controls gave a 6.1ng/uL average concentration, which means that our experiment had an efficiency of 6%. Further experiments will be needed in order to optimize the system.
NSTI Nanotech 2003 Conference Technical Program Abstract