Multichannel PCR-CE Microdevice for Genetic Analysis

Liu, Chung N.; Toriello, Nicholas M.; Mathies, Richard A.

doi:10.1021/ac060335k

Cited by 90 publications

(87 citation statements)

References 28 publications

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“…They were focused essentially on integrating the PCR amplification and the CE analysis using an optical LIF detection on a reusable multilayered glass microdevice [4] controlled and monitored by an enclosed electronic and optical system [5]. This approach uses different strategies to cleanup the PCR product and improve the sensitivity of the CE analysis [6,7] but requires the extraction and purification of the DNA analyte to be performed externally on the bench.…”

Section: Introductionmentioning

confidence: 99%

An automated instrument for human STR identification: Design, characterization, and experimental validation

Hurth¹,

Smith²,

Nordquist³

et al. 2010

Electrophoresis

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The microfluidic integration of an entire DNA analysis workflow on a fully integrated miniaturized instrument is reported using lab-on-a-chip automation to perform DNA fingerprinting compatible with CODIS standard relevant to the forensic community. The instrument aims to improve the cost, duration, and ease of use to perform a "sample-to-profile" analysis with no need for human intervention. The present publication describes the operation of the three major components of the system: the electronic control components, the microfluidic cartridge and CE microchip, and the optical excitation/detection module. Experimental details are given to characterize the level of performance, stability, reliability, accuracy, and sensitivity of the prototype system. A typical temperature profile from a PCR amplification process and an electropherogram of a commercial size standard (GeneScan 500™, Applied Biosystems) separation are shown to assess the relevance of the instrument to forensic applications. Finally, we present a profile from an automated integrated run where lysed cells from a buccal swab were introduced in the system and no further human intervention was required to complete the analysis.

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Section: Introductionmentioning

confidence: 99%

An automated instrument for human STR identification: Design, characterization, and experimental validation

Hurth¹,

Smith²,

Nordquist³

et al. 2010

Electrophoresis

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show abstract

“…7 Some reusable glass microfluidic devices must be equipped with removable, disposable valves because the PDMS valves would be destroyed by the harsh acid used to clean the device before reuse. 8,9 For these reasons, a large variety of interesting and useful chemistries may be unsuitable for use in native PDMS devices.…”

Section: Introductionmentioning

confidence: 99%

Teflon films for chemically-inert microfluidic valves and pumps

2008

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We present a simple method for fabricating chemically-inert Teflon microfluidic valves and pumps in glass microfluidic devices. These structures are modeled after monolithic membrane valves and pumps that utilize a featureless polydimethylsiloxane (PDMS) membrane sandwiched between two etched glass wafers. The limited chemical compatibility of PDMS has necessitated research into alternative materials for microfluidic devices. Previous work has shown that spincoated amorphous fluoropolymers and Teflon-fluoropolymer laminates can be fabricated and substituted for PDMS in monolithic membrane valves and pumps for space flight applications. However, the complex process for fabricating these spin-coated Teflon films and laminates may preclude their use in many research and manufacturing contexts. As an alternative, we show that commercially-available fluorinated ethylene-propylene (FEP) Teflon films can be used to fabricate chemically-inert monolithic membrane valves and pumps in glass microfluidic devices. The FEP Teflon valves and pumps presented here are simple to fabricate, function similarly to their PDMS counterparts, maintain their performance over extended use, and are resistant to virtually all chemicals. These structures should facilitate lab-on-a-chip research involving a vast array of chemistries that are incompatible with native PDMS microfluidic devices.

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“…Microchip electrophoresis has become one of the most powerful analytical tools in recent years [1][2][3] due to short analysis time, ease of integration, low consumption of samples and reagents, and feasibility for multichannel or highthroughput analysis. Gel electrophoresis (GE) on a microchip is gaining popularity in replacing conventional slab GE for the analysis of dsDNAs and proteins.…”

Section: Introductionmentioning

confidence: 99%

Integrated isotachophoretic stacking and gel electrophoresis on a plastic substrate and variations in detection dynamic range

2008

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In this study, we demonstrated an integrated ITP-gel electrophoresis (GE) device on a plastic substrate, in which 50 nL of samples could be hydrodynamically or electrokinetically injected and enriched by ITP into narrow bands and then subsequently introduced into a homogeneous GE channel for separation and detection. This microchip design rendered a simple introduction scheme for creating sandwiched stacking buffer system and flexibilities in choosing separation and stacking buffers independently. We used gel sieving buffers which compositions were different from those for stacking buffers to separate DNA and protein molecules based on sizing mechanism. Compared to conventional microchip GE, the sensitivity of microchip ITP-GE was estimated to increase by one to two orders of magnitude based on the dilution factor of the injected sample and the S/N ratio detected from the electropherogram. Moreover, it is interesting to note that ITP stacking leads to a preferential enhancement for analytes with lower concentrations compared to those with higher concentrations. Therefore, a reduction in the detection dynamic range for ITP-GE was gained. We demonstrated that ITP-GE could lead to 2-4-folds of reduction in the signal dynamic range for two PCR products in a mixture. Such advantage is demonstrated to be useful for the detection of two products amplified from a multiplex PCR in which one product is poorly amplified compared to the other.

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Multichannel PCR-CE Microdevice for Genetic Analysis

Cited by 90 publications

References 28 publications

An automated instrument for human STR identification: Design, characterization, and experimental validation

An automated instrument for human STR identification: Design, characterization, and experimental validation

Teflon films for chemically-inert microfluidic valves and pumps

Integrated isotachophoretic stacking and gel electrophoresis on a plastic substrate and variations in detection dynamic range

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