Purification of Nucleic Acids in Microfluidic Devices

Wen, Jian Kang; Legendre, Lindsay A.; Bienvenue, Joan M.; Landers, James P.

doi:10.1021/ac8014998

Cited by 152 publications

(135 citation statements)

References 56 publications

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“…Solid phase extraction (SPE) materials are widely used, in particular, highly cross-linked copolymers, functionalized copolymers, graphitized carbons, etc. (Boom et al 1990;Wen et al 2008). Among the solid phase sorbents, the most popular are magnetic particles containing immobilized ligands for capturing biological molecules (Berensmeier 2006;Gijs et al 2010).…”

Section: Introductionmentioning

confidence: 99%

Rotary-based platform with disposable fluidic modules for automated isolation of nucleic acids

Mamaev

Shaskolskiy

Dementieva

et al. 2015

Biomed Microdevices

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We describe the development and evaluation of a rotary-based platform with multiple disposable fluidic modules for simultaneous automatic nucleic acid (NA) isolation from up to 24 biological samples. The procedure is performed inside insulated individual disposable modules, which minimizes both the risk of infection of personnel and laboratory cross-contamination. Each module is a segment of a circular cylinder containing a leak-proof inlet port for sample input, reservoirs with lyophilized chemicals and solvents, fluidic channels, stoppers, valves, a waste reservoir and an outlet port equipped with the standard micro test tube for NA collection. The entire platform, apart from the rotor that accommodates 24 modules, consists of functional elements that provide spinning of the rotor, reagent mixing, pressure delivery, and heating of reaction mixtures. The transfer of the reaction mixtures inside the modules is performed either with rotation of the rotor or with excessive air pressure applied to the module's reservoirs. The entire process takes less than 40 min, starting from the sample loading to the recovery of the purified NA, and it allows NA isolation both from bacterial cells and viral particles. The feasibility and reproducibility of the developed platform was demonstrated by the NA isolation from suspensions of Bacillus thuringiensis and Mycobacterium tuberculosis cells within a concentration range of 10(8) to 10(2) cells/ml. Isolation of NAs from blood plasma samples with varying concentration of hepatitis B and C viruses from 10(7) to 10(2) particles/ml were also successful. The purity and integrity of the extracted NAs were both reliable for performing quantitative PCR.

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Section: Introductionmentioning

confidence: 99%

Rotary-based platform with disposable fluidic modules for automated isolation of nucleic acids

Mamaev

Shaskolskiy

Dementieva

et al. 2015

Biomed Microdevices

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“…1 Extending the surface area is a key aspect in many applications like separations, 2 extraction, 3 purification of both small molecules and macromolecules, 4 filtration of bioparticles, 5 and immobilization of enzymes, antibodies or cells. 6 With the advent of microfluidics, monoliths were implemented in microchannels, providing an easily fabricated and versatile support needed in many research areas.…”

Section: Introductionmentioning

confidence: 99%

Advances in monoliths and related porous materials for microfluidics

2016

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In recent years, the use of monolithic porous polymers has seen significant growth. These materials present a highly useful support for various analytical and biochemical applications. Since their introduction, various approaches have been introduced to produce monoliths in a broad range of materials. Simple preparation has enabled their easy implementation in microchannels, extending the range of applications where microfluidics can be successfully utilized. This review summarizes progress regarding monoliths and related porous materials in the field of microfluidics between 2010 and 2015. Recent developments in monolith preparation, solid-phase extraction, separations, and catalysis are critically discussed. Finally, a brief overview of the use of these porous materials for analysis of subcellular and larger structures is given.

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“…Cepheid GeneXpert, Biocartis and. Abbott i-ST AT ), many of these systems still lack commercial maturit y (Lazcka et al 2007), especially when they include sample preparation steps (Wen et al 2008). They further often require expensive hardware for optical readout or fluidic control and are usable for a small range of applications only (Yager et al 2008).…”

Section: Introductionmentioning

confidence: 99%

Centrifugal LabTube platform for fully automated DNA purification and LAMP amplification based on an integrated, low-cost heating system

Hoehl

Weißert

Dannenberg

et al. 2014

Biomed Microdevices

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Abstract:This paper introduces a disposable battery-driven heating system for loop-mediated isothermal DNA amplification (LAMP) inside a centrifugally-driven DNA p urification platform (LabT ube). We demonstrate LabT ube-based fully automated DNA purification of as low as 100 cell-equivalents of verotoxin-producing Escherichia coli (VT EC) in water, milk and apple juice in a laboratory centrifuge, followed by integrated and automated LAMP amplification with a reduction of hands-on time from 45 to 1min. The heating system consists of two parallel SMD thick film resistors and a NT C as heating and temperature sensing elements. They are driven by a 3V battery and controlled by a microcontroller. The LAMP reagents are stored in the elution chamber and the amplification starts immediately after the eluate is purged into the chamber. The LabT ube, including a microcontroller-based heating system, demonstrates contamination-free and automated sample-to-answer nucleic acid testing within a laboratory centrifuge. The heating system can be easily parallelized within one LabT ube and it is deployable for a variety of heating and electrical applications.

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Purification of Nucleic Acids in Microfluidic Devices

Abstract: The functionality of micropillars, microposts, silica beads, silica particles, sol-gels, and porous monoliths provides a framework for sample preparation and analysis for an integrated microfluidic system.

Cited by 152 publications

References 56 publications

Rotary-based platform with disposable fluidic modules for automated isolation of nucleic acids

Rotary-based platform with disposable fluidic modules for automated isolation of nucleic acids

Advances in monoliths and related porous materials for microfluidics

Centrifugal LabTube platform for fully automated DNA purification and LAMP amplification based on an integrated, low-cost heating system

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