Microchip-based Real-time PCR
Polymerase chain reaction (PCR) is the most widely used method in molecular biology area. PCR process exponentially amplifies DNA via enzymatic replication, capable of producing millions of copies of a DNA fragment from just a few copies. This effect is achieved by sequential heating and cooling of the analyzed sample with reagents and detecting amplified products of the reaction if specific targets are present. Test system reagents define target pieces of DNA to be amplified and also the level of amplification of the products.
Classical PCR thermocycler instruments using plastic tubes, well-plates or glass capillary have shortcomings, limiting further technology development opportunities. These well-known limits for PCR advancement can be summarized as follows: low rates of sample heating and cooling resulting in longer analysis time; high temperature gradient in a sample during heating leading to low reaction specificity; high volumes of reagents consumed (5-50 mL) leading to higher cost per test; labour-intensive test-preparation stage resulting in higher labour costs per test; freeze-chain storage and transportation conditions for the reagents.
To improve the PCR method and instrument efficiency researchers turned to microfluidics technology and its applications for PCR analysis systems. Microfluidics is an area of so-called lab-on-chip systems that use photolithographic technology from microelectronics for developing miniaturized systems for analytical testing of chemical substances. Main advantages of the microfluidic chips for PCR applications are fast analysis, low labor and reagents consumption and convenient process of PCR analysis and reagents storage.
Lumex Instruments focused its research efforts on open-well microfluidic chips, providing all the benefits of microfluidic technology operating in simple PCR thermocycler. As a result of the efforts Lumex created a compact PCR thermocycler AriaDNA that uses simple optical scheme with minimum moving parts, making system almost maintenance free. We have also developed a technology for mass production of the microchips. Application of high heat conductivity materials for microchips and modern technological processes for protective layers deposition allow to reach long storage life of the PCR reagents on the microchips at room temperature conditions. Microchips with reagents are designed for the specific user applications in the areas of medicine, veterinary and agriculture.
