Chip PCR. I. Surface passivation of microfabricated silicon-glass chips for PCR

Shoffner, Mann A.; Cheng, Jing; Hvichia, Georgi E.; Kricka, Larry J.; Wilding, Peter

doi:10.1093/nar/24.2.375

Cited by 253 publications

(176 citation statements)

References 17 publications

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“…1B (the right diagram). 7,12 Besides surface passivation of the silicon-side reaction cham-ber, attention should also be given to the surface chemistry of the glass substrate within the reaction chamber. Properties of both glass surfaces (Corning 7740 glass and MPTS-modified ITO glass) in our PCR-EC microdevices are compatible with PCR reaction.…”

Section: Resultsmentioning

confidence: 99%

Microfabricated PCR-electrochemical device for simultaneous DNA amplification and detection

2003

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Microfabricated silicon/glass-based devices with functionalities of simultaneous polymerase chain reaction (PCR) target amplification and sequence-specific electrochemical (EC) detection have been successfully developed. The microchip-based device has a reaction chamber (volume of 8 µl) formed in a silicon substrate sealed by bonding to a glass substrate. Electrode materials such as gold and indium tin oxide (ITO) were patterned on the glass substrate and served as EC detection platforms where DNA probes were immobilized. Platinum temperature sensors and heaters were patterned on top of the silicon substrate for real-time, precise and rapid thermal cycling of the reaction chamber as well as for efficient target amplification by PCR. DNA analyses in the integrated PCR-EC microchip start with the asymmetric PCR amplification to produce single-stranded target amplicons, followed by immediate sequence-specific recognition of the PCR product as they hybridize to the probe-modified electrode. Two electrochemistry-based detection techniques including metal complex intercalators and nanogold particles are employed in the microdevice to achieve a sensitive detection of target DNA analytes. With the integrated PCR-EC microdevice, the detection of trace amounts of target DNA (as few as several hundred copies) is demonstrated. The ability to perform DNA amplification and EC sequence-specific product detection simultaneously in a single reaction chamber is a great leap towards the realization of a truly portable and integrated DNA analysis system.

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Section: Resultsmentioning

confidence: 99%

Microfabricated PCR-electrochemical device for simultaneous DNA amplification and detection

2003

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“…To minimize the surface affinity for molecules from the solution, we over-coated micro-wells with an additional silicon oxide dielectric layer 200 nm thick, by magnetron sputtering [24][25][26], forming titanium-silicon oxide coated and aluminum-silicon oxide coated fiber-optic slides. The surface morphologies of the four different coatings were examined by atomic force microscopy (AFM).…”

Section: Surface Coatingmentioning

confidence: 99%

Improved picoliter-sized micro-reactors for high-throughput biological analysis

Han

Yuan

Wei

et al. 2013

Sci. China Life Sci.

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High-throughput pyrosequencing, carried out in millions of picoliter-sized reactors on a fiber-optic slide, is known for its longer read length. However, both optical crosstalk (which reduces the signal-to-noise ratio of CCD images) and chemical retention adversely affect the accuracy of chemiluminescence determination, and ultimately decrease the read length and the accuracy of pyrosequencing results. In this study, both titanium and oxidized aluminum films were deposited on the side walls and upper faces of micro-reactor slides to enhance optical isolation; the films reduced the inter-well crosstalk by one order of magnitude. Subsequently, chemical retention was shown to be caused by the lower diffusion coefficient of the side walls of the picolitersized reactors because of surface roughness and random pores. Optically isolated fiber-optic slides over-coated with silicon oxide showed smoother surface morphology, resulting in little chemical retention; this was further confirmed with theoretical calculations. Picoliter-sized micro-reactors coated with titanium-silicon oxide films showed the least inter-well optical crosstalk and chemical retention; these properties are expected to greatly improve the high-throughput pyrosequencing performance.high throughput analysis, picoliter-sized micro-reactor, surface coating Citation:Han W J, Yuan L N, Wei Q Q, et al. Improved picoliter-sized micro-reactors for high-throughput biological analysis.

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“…As reaction chambers are miniaturized, the surface-to-volume ratio increases. Previous workers have focused on surface coatings to improve reaction performance (17). We have successfully performed many enzymatic reactions in polymeric cartridge reactors with 5-30 µl volumes (Table 2) in which some reformulation of reagents combined with a parylene surface coating proved successful.…”

Section: Integration Of Multiple Enzymatic Reactions With Reagent Hanmentioning

confidence: 99%

A miniature integrated device for automated multistep genetic assays

Anderson¹,

Su²,

Bogdan³

et al. 2000

Nucleic Acids Research

204

101

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A highly integrated monolithic device was developed that automatically carries out a complex series of molecular processes on multiple samples. The device is capable of extracting and concentrating nucleic acids from milliliter aqueous samples and performing microliter chemical amplification, serial enzymatic reactions, metering, mixing and nucleic acid hybridization. The device, which is smaller than a credit card, can manipulate over 10 reagents in more than 60 sequential operations and was tested for the detection of mutations in a 1.6 kb region of the HIV genome from serum samples containing as few as 500 copies of the RNA. The elements in this device are readily linked into complex, flexible and highly parallel analysis networks for high throughput sample preparation or, conversely, for low cost portable DNA analysis instruments in point-of-care medical diagnostics, environmental testing and defensive biological agent detection.

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Chip PCR. I. Surface passivation of microfabricated silicon-glass chips for PCR

Cited by 253 publications

References 17 publications

Microfabricated PCR-electrochemical device for simultaneous DNA amplification and detection

Microfabricated PCR-electrochemical device for simultaneous DNA amplification and detection

Improved picoliter-sized micro-reactors for high-throughput biological analysis

A miniature integrated device for automated multistep genetic assays

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