A rapid DNA digestion system.
This paper presents a novel microfluidic DNA digestion system incorporating a high performance micro-mixer. Through the appropriate control of fixed and periodic switching DC electric fields, electrokinetic forces are established to mix the DNA and restriction enzyme samples and to drive them through the reaction column of the device. The experimental and numerical results show that a mixing performance of 98% can be achieved within a mixing channel of length 1.6 mm when a 150 V/cm driving voltage and a 5 Hz switching frequency are applied. The relationship between the mixing performance, switching frequency, and main applied electric field is derived. It is found that the optimal switching frequency depends upon the magnitude of the main applied electric field. The successful digestion of lambda-DNA using Eco RI restriction enzyme is demonstrated. The DNA-enzyme reaction is completed within 15 min in the proposed microfluidic system, compared to 50 min in a conventional large-scale system. Hence, the current device provides a valuable tool for rapid lambda-DNA digestion, while its mixer system delivers a simple yet effective solution for mixing problems in the micro-total-analysis-systems field.
Fu
Lung-Ming
LM
Department of Materials Engineering, National Pingtung University of Science and Technology, Pingtung, Taiwan, 912, China.
Lin
Che-Hsin
CH
D010582:Bacteriophage lambda / Q000187:drug effects*; D004262:DNA Restriction Enzymes / Q000737:chemistry*; D004279:DNA, Viral / Q000737:chemistry* / Q000302:isolation & purification*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D017022:Flow Injection Analysis / Q000295:instrumentation* / Q000379:methods; D044085:Microfluidics / Q000295:instrumentation* / Q000379:methods
D004279:DNA, Viral; D004262:DNA Restriction Enzymes