Developments toward a complete micro-total analysis system for Duchenne muscular dystrophy diagnosis

Ferrance, Jerome P.; Wu, Qirong; Giordano, Braden C.; Hernandez, Casandra; Kwok, Yien Chian; Snow, Karen; Thibodeau, S. N.; Landers, James P.

doi:10.1016/j.aca.2003.08.067

Cited by 75 publications

(52 citation statements)

References 23 publications

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“…In this system, post-amplification PCR product analysis was performed off-chip. IR-mediated PCR has since been integrated with microchip CE for the analysis of ␤-globin in a genomic DNA sample (34). Another order-of-magnitude reduction in PCR chamber volume was achieved on a monolithic micromachined glass substrate (35; Figure 2d).…”

Section: Sample Preparationmentioning

confidence: 99%

Microfluidic Systems for Integrated, High-Throughput DNA Analysis

Kelly¹,

Woolley²

2005

Anal. Chem.

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B ecause DNA is the blueprint for life in all its forms, it is no wonder that nucleic acids are of such interest to scientists. The Human Genome Project, completed in 2003, produced a wealth of information, including the number and average size of human genes, the fraction of the genome that codes for proteins, and the degree of sequence similarity, both among humans and compared with other organisms (1, 2). More importantly, deciphering the human genome should allow researchers to unravel the complex interplay between genetic and environmental factors involved in many diseases, and this should facilitate the development of therapeutic agents (1).The abundance of genomic information has been made possible in large part by improved analytical methods for nucleic acids. If all of the sequencing for the Human Genome Project had been performed with the slab gel technology that was available when the project began, the raw data would have been obtained at a rate of ~600 bases/h (3). The electrophoretic separations for just onefold coverage of the human genome would have taken millions of instrument-hours, in addition to the time required for sample preparation and data analysis. Development of capillary array electrophoresis (CAE), in which multiple samples are electrophoretically analyzed in parallel in a bundle of ~100-µm-diam capillaries, was a major breakthrough in DNA analysis (4 -7 ). High-throughput instruments with parallel capillaries for electro-

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Section: Sample Preparationmentioning

confidence: 99%

Microfluidic Systems for Integrated, High-Throughput DNA Analysis

Kelly¹,

Woolley²

2005

Anal. Chem.

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“…Since the first report concerning continuous-flow PCR by Kopp in 1998, 7 microfluidic PCR devices performing all steps of PCR on a single chip have been anticipated. [8][9][10] A typical device is composed of three temperature zones (denaturation, annealing and extension) and a long serpentine microchannel that carries the PCR reagents to each temperature zone. This continuous-flow PCR technique can perform rapid thermal cycles because the element related to heat capacity is the PCR solution itself, and is not dependent on the heating-and-cooling ramping rates of the reactor vessels or the temperature-controlled metal blocks.…”

Section: Introductionmentioning

confidence: 99%

Rapid and Highly Sensitive Detection by a Real-time Polymerase Chain Reaction Using a Chip Coated with Its Reagents

et al. 2014

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On-site detection by flow-through polymerase chain reaction (PCR) microfluidic systems for rapid and highly sensitive analysis, are significantly desired for bioanalytical and medical research. The conventional continuous-flow PCR chips realized rapid detection, but their sensitivity was very low (10 6 to 10 8 copies μL). We improved this drawback by coating the chip with a PCR reagents mixture, and succeed to obtain a rapid and highly sensitive detection by using a segment-flow PCR system. In the present work, we developed a portable segment-flow PCR system for practical use. PCR was performed for the uid A gene in E. coli. By real-time segment-flow PCR using coated chips, we realized rapid detection in 8 min and a high sensitivity of 4 cells μL -1 . The sensitivity by the segment-flow PCR chip was the same as that of a conventional thermal cycler. Moreover, the detection speed of our segment-flow PCR chip was 15-times as rapid as that of the conventional thermal cycler.

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“…[2][3][4] In principle, PCR reagents would be loaded into long micro-flow channels and repeatedly carried to individual temperature zones for denaturation, extension and annealing. This flow-through technique would achieve PCR in a short time period because it is independent of the thermal conductivity and ramping rate for heating and cooling.…”

Section: Introductionmentioning

confidence: 99%

A Practical Liquid Plug Flow-through Polymerase Chain-Reaction System Based on a Heat-Resistant Resin Chip

et al. 2011

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Flow-through polymerase chain reaction (PCR) microfluidic systems for fast, small-volume DNA amplification on a single chip are significantly impacting medical and bioanalytical research. We have fabricated an improved, practical flow-through PCR chip by weighting a pressure-sensitive polyolefin (PSP) film onto a cyclo-olefin polymer (COP) substrate. The substrate was cut so as to produce microchannels, and was used to amplify DNA using a small moving liquid plug, in contrast to conventional continuous-flow-through PCR. Infrared (IR) imaging-based thermal analysis of the PSP film enabled the gradient and uniformity of the rapidly flowing microfluid to be accurately visualized, because the PSP film is thin, transparent and heat resistant. The liquid plug flow-through PCR, operating at 150 μl min -1 , showed approximately 55% amplification compared to a commercial PCR instrument, and required only 250 s for 40 cycles. To our knowledge, this is the fastest cycling time reported to date. In a real test sample, the liquid plug flow-through PCR could detect the presence or absence of anthrax in a sample solution in approximately 250 s. Thus, this liquid plug flow-through PCR system, based on our novel PCR chip, excelled in a practical applications compared to other devices.

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Developments toward a complete micro-total analysis system for Duchenne muscular dystrophy diagnosis

Cited by 75 publications

References 23 publications

Microfluidic Systems for Integrated, High-Throughput DNA Analysis

Microfluidic Systems for Integrated, High-Throughput DNA Analysis

Rapid and Highly Sensitive Detection by a Real-time Polymerase Chain Reaction Using a Chip Coated with Its Reagents

A Practical Liquid Plug Flow-through Polymerase Chain-Reaction System Based on a Heat-Resistant Resin Chip

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