A modular microfluidic system for deoxyribonucleic acid identification by short tandem repeat analysis.

Microfluidic technology has been utilized in the development of a modular system for DNA identification through STR (short tandem repeat) analysis, reducing the total analysis time from the ∼6 h required with conventional approaches to less than 3h. Results demonstrate the utilization of microfluidic devices for the purification, amplification, separation and detection of 9 loci associated with a commercially-available miniSTR amplification kit commonly used in the forensic community. First, DNA from buccal swabs purified in a microdevice was proven amplifiable for the 9 miniSTR loci via infrared (IR)-mediated PCR (polymerase chain reaction) on a microdevice. Microchip electrophoresis (ME) was then demonstrated as an effective method for the separation and detection of the chip-purified and chip-amplified DNA with results equivalent to those obtained using conventional separation methods on an ABI 310 Genetic Analyzer. The 3-chip system presented here demonstrates development of a modular, microfluidic system for STR analysis, allowing for user-discretion as to how to proceed after each process during the analysis of forensic casework samples.

[1]  John M. Butler,et al.  Forensic DNA Typing: Biology, Technology, and Genetics of STR Markers , 2001 .

[2]  James P Landers,et al.  AOTF-based multicolor fluorescence detection for short tandem repeat (STR) analysis in an electrophoretic microdevice. , 2008, Lab on a chip.

[3]  Steven A Soper,et al.  High resolution DNA separations using microchip electrophoresis. , 2007, Journal of separation science.

[4]  Jerome P Ferrance,et al.  Chitosan-coated silica as a solid phase for RNA purification in a microfluidic device. , 2009, Analytical chemistry.

[5]  Daniel J Marchiarullo,et al.  Pressure injection on a valved microdevice for electrophoretic analysis of submicroliter samples. , 2005, Analytical chemistry.

[6]  William A. Mcmillan,et al.  Rapid, automated nucleic acid probe assays using silicon microstructures for nucleic acid concentration. , 1999, Journal of biomechanical engineering.

[7]  James P Landers,et al.  Infrared temperature control system for a completely noncontact polymerase chain reaction in microfluidic chips. , 2007, Analytical chemistry.

[8]  S. Jacobson,et al.  Integrated system for rapid PCR-based DNA analysis in microfluidic devices. , 2000, Analytical chemistry.

[9]  Andreas Manz,et al.  Micromachining of monocrystalline silicon and glass for chemical analysis systems A look into next century's technology or just a fashionable craze? , 1991 .

[10]  A. Barron,et al.  Hydrophobically modified polyacrylamide block copolymers for fast, high‐resolution DNA sequencing in microfluidic chips , 2008, Electrophoresis.

[11]  S N Austad.  Forensic DNA typing. , 1992, Science.

[12]  A. Barron,et al.  Poly(acrylamide-co-alkylacrylamides) for electrophoretic DNA purification in microchannels. , 2005, Analytical chemistry.

[13]  James P. Landers,et al.  Toward a Simplified Microfluidic Device for Ultra-Fast Genetic Analysis with Sample-In/Answer-Out Capability: Application to T-Cell Lymphoma Diagnosis , 2008 .

[14]  Jerome P Ferrance,et al.  Microchip‐Based Cell Lysis and DNA Extraction from Sperm Cells for Application to Forensic Analysis , 2006, Journal of forensic sciences.

[15]  J. Landers,et al.  Volume reduction solid phase extraction of DNA from dilute, large-volume biological samples. , 2010, Forensic science international. Genetics.

[16]  Jerome P Ferrance,et al.  Extrinsic Fabry-Perot interferometry for noncontact temperature control of nanoliter-volume enzymatic reactions in glass microchips. , 2005, Analytical chemistry.

[17]  James P Landers,et al.  Advances in polymerase chain reaction on microfluidic chips. , 2005, Analytical chemistry.

[18]  Jerome P Ferrance,et al.  A simple, valveless microfluidic sample preparation device for extraction and amplification of DNA from nanoliter-volume samples. , 2006, Analytical chemistry.

[19]  Annelise E Barron,et al.  Polymer systems designed specifically for DNA sequencing by microchip electrophoresis: A comparison with commercially available materials , 2008, Electrophoresis.

[20]  Michael G. Roper,et al.  A fully integrated microfluidic genetic analysis system with sample-in–answer-out capability , 2006, Proceedings of the National Academy of Sciences.

[21]  Lindsay A. Legendre,et al.  Rapid DNA amplification in glass microdevices. , 2006, Methods in molecular biology.

[22]  K. A. Wolfe,et al.  Microchip-based purification of DNA from biological samples. , 2003, Analytical chemistry.

[23]  Gillian Tully,et al.  Integrated microfluidic system for rapid forensic DNA analysis: sample collection to DNA profile. , 2010, Analytical chemistry.

[24]  John M. Butler,et al.  Forensic DNA typing : biology & technology behind STR markers , 2001 .

[25]  Richard A Mathies,et al.  Microfabricated bioprocessor for integrated nanoliter-scale Sanger DNA sequencing. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[26]  James P Landers,et al.  Multicolor fluorescence detection on an electrophoretic microdevice using an acoustooptic tunable filter. , 2006, Analytical chemistry.

[27]  Richard A Mathies,et al.  A Forensic Laboratory Tests the Berkeley Microfabricated Capillary Array Electrophoresis Device * , 2008, Journal of forensic sciences.

[28]  Brian McKenna,et al.  A high‐performance multilane microdevice system designed for the DNA forensics laboratory , 2004, Electrophoresis.

[29]  A. Lehninger.  Principles of Biochemistry , 1984 .

[30]  J. Landers,et al.  Evaluation of silica resins for direct and efficient extraction of DNA from complex biological matrices in a miniaturized format. , 2000, Analytical biochemistry.