On-line cell lysis and DNA extraction on a microfluidic biochip fabricated by microelectromechanical system technology.
Integrating cell lysis and DNA purification process into a micrototal analytical system (microTAS) is one critical step for the analysis of nucleic acids. On-chip cell lysis based on a chemical method is realized by sufficient blend of blood sample and the lyzing reagent. In this paper two mixing models, T-type mixing model and sandwich-type mixing model, are proposed and simulation of those models is conducted. Result of simulation shows that the sandwich-type mixing model with coiled channel performs best and this model is further used to construct the microfluidic biochip for on-line cell lysis and DNA extraction. The result of simulation is further verified by experiments. It asserts that more than 80% mixing of blood sample and lyzing reagent which guarantees that completed cell lysis can be achieved near the inlet location when the cell/buffer velocity ratio is less than 1:5. After cell lysis, DNA extraction by means of a solid-phase method is implemented by using porous silicon matrix which is integrated in the biochip. During continuous flow process in the microchip, rapid cell lysis and PCR-amplifiable genomic DNA purification can be achieved within 20 min. The potential of this microfluidic biochip is illustrated by pretreating a whole blood sample, which shows the possibility of integration of sample preparation, PCR, and separation on a single device to work as portable point-of-care medical diagnostic system.
Chen
Xing
X
State Key Laboratory of Transducer Technology, Institute of Electronics, Chinese Academy of Sciences, Beijing, P. R. China. chenxing2004star@yahoo.com.cn
Cui
Da Fu
DF
Liu
Chang Chun
CC
D000818:Animals; D052160:Benzothiazoles; D001773:Blood Cells / Q000737:chemistry*; D002021:Buffers; D004247:DNA / Q000097:blood / Q000302:isolation & purification*; D003959:Diamines; D004563:Electrochemistry; D004867:Equipment Design; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods; D005456:Fluorescent Dyes; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D008956:Models, Chemical; D009930:Organic Chemicals; D016133:Polymerase Chain Reaction; D016062:Porosity; D011804:Quinolines; D051381:Rats; D012825:Silicon
D052160:Benzothiazoles; D002021:Buffers; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I; D004247:DNA; D012825:Silicon