Microfluidic chip for fast nucleic acid hybridization.

The design and experimental verification of a fast nucleic acid hybridization microchip using the fluidic velocity and strain rate effects was conducted. This hybridization chip was able to increase the hybridization signal 6-fold, reduce non-specific target-probe binding and background noise within 30 min, as compared to conventional hybridization methods, which may take from 4 h to overnight. Excellent correlation between experimental results and simulation analysis was obtained in this study. A detailed study of a newly designed microfluidic chip for enhancing hybridization was conducted. Three different designs of devices were fabricated and tested. Two different lengths of targets, 25-mer oligonucleotide and 1.4 kb ssDNA, were tested in this study. The hybridization efficiency can be improved by introducing velocity and extensional strain rate to the sample. This study demonstrates that the signal in the proposed method exhibits intensities 6-fold higher than those in static conditions. The necessary time for the hybridization process can be reduced from overnight to 30 min using the methods developed in this study. Experimental results also show that the strain rate provides stronger effect on hybridization than that of velocity. Combining hybridization with microfluidic concepts of velocity and strain rate effects may provide additional specificity and efficiency in nucleic acid detection and genomic study. This microfluidic hybridization chip can provide potential application in genomic study.

[1]  A. Mirzabekov,et al.  Theoretical analysis of the kinetics of DNA hybridization with gel-immobilized oligonucleotides. , 1996, Biophysical journal.

[2]  Dorian Liepmann,et al.  Effect of Flow on Complex Biological Macromolecules in Microfluidic Devices , 2001 .

[3]  Philip LeDuc,et al.  Dynamics of individual flexible polymers in a shear flow , 1999, Nature.

[4]  B. J. Cheek,et al.  Chemiluminescence detection for hybridization assays on the flow-thru chip, a three-dimensional microchannel biochip. , 2001, Analytical chemistry.

[5]  Yu-Cheng Lin,et al.  Rapid micro-PCR system for hepatitis C virus amplification , 2000, SPIE MOEMS-MEMS.

[6]  Chong-Ching Chang,et al.  Experimental Verification of A Bi-Directional Driving System for Microfluids , 2002 .

[7]  Ronald W. Davis,et al.  Quantitative Monitoring of Gene Expression Patterns with a Complementary DNA Microarray , 1995, Science.

[8]  M. Heller,et al.  Rapid determination of single base mismatch mutations in DNA hybrids by direct electric field control. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[9]  R. G. Larsona,et al.  Brownian dynamics simulations of a DNA molecule in an extensional flow field , 1999 .

[10]  D E Smith,et al.  Single polymer dynamics in an elongational flow. , 1997, Science.

[11]  S. Edwards,et al.  The Theory of Polymer Dynamics , 1986 .

[12]  G. McKinley,et al.  Stress relaxation and elastic decohesion of viscoelastic polymer solutions in extensional flow , 1996 .

[13]  J. Nieuwkoop,et al.  Elongation and subsequent relaxation measurements on dilute polyisobutylene solutions , 1996 .

[14]  Ming-Yuan Huang,et al.  A rapid micro-polymerase chain reaction system for hepatitis C virus amplification , 2000 .

[15]  B. Zimm,et al.  Fracture of polymer chains in extensional flow : experiments with DNA, and a molecular-dynamics simulation , 1990 .

[16]  Douglas E. Smith,et al.  Single-polymer dynamics in steady shear flow. , 1999, Science.

[17]  J P Landers,et al.  Noncontact infrared-mediated thermocycling for effective polymerase chain reaction amplification of DNA in nanoliter volumes. , 2000, Analytical chemistry.

[18]  Gareth H. McKinley,et al.  Relaxation of dilute polymer solutions following extensional flow 1 Dedicated to the memory of Profe , 1998 .

[19]  S. P. Fodor,et al.  Light-generated oligonucleotide arrays for rapid DNA sequence analysis. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[20]  Hye Jung Cho,et al.  Precise temperature control and rapid thermal cycling in a micromachined DNA polymerase chain reaction chip , 2002 .

[21]  L. Hood,et al.  DNA sequence determination by hybridization: a strategy for efficient large-scale sequencing. , 1993, Science.

[22]  S. Mangru,et al.  Dynamic DNA hybridization on a chip using paramagnetic beads. , 1999, Analytical chemistry.

[23]  Ronald G. Larson,et al.  Brownian dynamics simulations of single DNA molecules in shear flow , 2000 .

[24]  Robin H. Liu,et al.  Plastic biochannel hybridization devices: a new concept for microfluidic DNA arrays. , 2002, Analytical biochemistry.

[25]  R. Larson,et al.  Brownian dynamics simulations of a DNA molecule in an extensional flow field , 1999 .

[26]  Ming-Yuan Huang,et al.  Simulation and experimental validation of micro polymerase chain reaction chips , 2000 .

[27]  James L. Winkler,et al.  Accessing Genetic Information with High-Density DNA Arrays , 1996, Science.

[28]  M. Heller,et al.  Electric field directed nucleic acid hybridization on microchips. , 1997, Nucleic acids research.

[29]  R. Reich,et al.  Direct detection of nucleic acid hybridization on the surface of a charge coupled device. , 1994, Nucleic acids research.

[30]  B. Stillman,et al.  Expression microarray hybridization kinetics depend on length of the immobilized DNA but are independent of immobilization substrate. , 2001, Analytical biochemistry.

[31]  D. Garling,et al.  Rapid cycle DNA amplification: time and temperature optimization. , 1991, BioTechniques.

[32]  F. White Viscous Fluid Flow , 1974 .

[33]  Stewart J. Fallon,et al.  Rapid, high fidelity analysis of simple sequence repeats on an electronically active DNA microchip. , 2000, Nucleic acids research.

[34]  K. Shimizu,et al.  Response of Flexible Polymers to a Sudden Elongational Flow , 1998 .

[35]  P. Brown,et al.  Yeast microarrays for genome wide parallel genetic and gene expression analysis. , 1997, Proceedings of the National Academy of Sciences of the United States of America.