PCR microfluidic devices for DNA amplification

Zhang, Chunsun; Xu, Jie; Ma, Wenxue; Wen-ling, Zheng

doi:10.1016/j.biotechadv.2005.10.002

Cited by 526 publications

(372 citation statements)

References 173 publications

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“…Despite growing interest in developing "lab-ona-chip" devices for inexpensive DNA isolation [2,3] [6] the utility of readily available, unmodified glass surfaces in DNA capture chambers has not been described. Several devices have been designed to miniaturize the sample sizes for micro-PCR analysis [7], and silica based DNA capture surfaces have been engineered with higher binding capacity. For example, silica resins have been optimized for DNA capture in microchambers in the presence of chaotropic salts [8,9].…”

Section: Discussionmentioning

confidence: 99%

Capture of genomic DNA on glass microscope slides

Nanassy

Haydock

Reed

2007

Analytical Biochemistry

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It is well known that DNA strands bind to silica surfaces in the presence of high concentrations of chaotropic salts. We developed simple methods to evaluate binding and recovery of DNA on flat glass microscope slides and compared their properties to commercially available silica "spin columns". Surprisingly, genomic DNA was recovered efficiently from untreated glass slides. Binding and elution times from glass slides were optimized in experiments with DNA samples of various sizes and defined buffers. Average DNA recovery from 500 ng of input genomic DNA varied from 25-53 % for the glass slide protocol. Yields were comparable to spin columns. Human serum albumin and plasma components decreased DNA binding to glass several-fold in a concentration dependent manner. These results support the concept of using flat glass slides as DNA purification surfaces in microfluidics devices for PCR sample preparation. Keywords DNA extraction; silica surfaces; PCR; microfluidicsThe use of powdered glass or other high surface area silica materials has been widely adopted as a basic technique for purifying DNA from biological samples [1]. DNA strands bind efficiently to silica particles in the presence of high concentrations of chaotropic salts such as guanidinium thiocyanate or sodium perchlorate. After washing away contaminating substances with high salt buffers, the bound DNA is released from the silica surface in the presence of water or low salt buffer. The recovered DNA is suitable for use in PCR or other amplification methods. These methods are used extensively in medical diagnostics and forensics, in recombinant technology and for molecular genetics.Silica-based DNA purification is compatible with microfluidic diagnostic devices. Such "lab on a chip" devices may help bring the benefits of PCR-based molecular diagnosis to users and point-of-care settings that lack the capacity for standard laboratory-based DNA purification [2] [3]. In such applications there are significant pressures to keep costs low. Therefore, DNA purification devices must be designed to use the least expensive materials possible, and without unnecessary engineering. Early researchers showed that the curved inner surfaces of glass test tubes could be used to purify DNA from agarose gels, but their binding capacity was deemed too small for most preparative experiments [4]. Currently, it is standard practice to maximize DNA binding surface area, for example by using silica glass bead matrices. However, such engineering measures may not be necessary to achieve satisfactory DNA purification yields.

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Section: Discussionmentioning

confidence: 99%

Capture of genomic DNA on glass microscope slides

Nanassy

Haydock

Reed

2007

Analytical Biochemistry

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“…In the past decade, microfluidic system, also called "lab-on-chip (LOC)", "bio-chip", or "micro-total-analysis-system (μTAS)", has attracted attention because of its capability of combining engineering and life science [1][2][3]. Therefore, it is often interpreted as a miniaturized and automatic version of a conventional laboratory.…”

Section: Introductionmentioning

confidence: 99%

Review on Impedance Detection of Cellular Responses in Micro/Nano Environment

Lei

2014

Micromachines

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Abstract:In general, cell culture-based assays, investigations of cell number, viability, and metabolic activities during culture periods, are commonly performed to study the cellular responses under various culture conditions explored. Quantification of cell numbers can provide the information of cell proliferation. Cell viability study can understand the percentage of cell death under a specific tested substance. Monitoring of the metabolic activities is an important index for the study of cell physiology. Based on the development of microfluidic technology, microfluidic systems incorporated with impedance measurement technique, have been reported as a new analytical approach for cell culture-based assays. The aim of this article is to review recent developments on the impedance detection of cellular responses in micro/nano environment. These techniques provide an effective and efficient technique for cell culture-based assays.

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“…This implies high cost when manufacturing and energy waste during high speed heating and cooling. In this decade, microfluidics technology has developed rapidly and has been applied to the field of PCR [3][4][5][6][7][8]. This platform pumps a low volume of PCR buffer to pass through two or three constant-temperature zones repeatedly.…”

Section: Introductionmentioning

confidence: 99%

Development of Capillary Loop Convective Polymerase Chain Reaction Platform with Real-Time Fluorescence Detection

et al. 2017

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Polymerase chain reaction (PCR) has been one of the principal techniques of molecular biology and diagnosis for decades. Conventional PCR platforms, which work by rapidly heating and cooling the whole vessel, need complicated hardware designs, and cause energy waste and high cost. On the other hand, partial heating on the various locations of vessels to induce convective solution flows by buoyancy have been used for DNA amplification in recent years. In this research, we develop a new convective PCR platform, capillary loop convective polymerase chain reaction (clcPCR), which can generate one direction flow and make the PCR reaction more stable. The U-shaped loop capillaries with 1.6 mm inner diameter are designed as PCR reagent containers. The clcPCR platform utilizes one isothermal heater for heating the bottom of the loop capillary and a CCD device for detecting real-time amplifying fluorescence signals. The stable flow was generated in the U-shaped container and the amplification process could be finished in 25 min. Our experiments with different initial concentrations of DNA templates demonstrate that clcPCR can be applied for precise quantification. Multiple sample testing and real-time quantification will be achieved in future studies.

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PCR microfluidic devices for DNA amplification

Cited by 526 publications

References 173 publications

Capture of genomic DNA on glass microscope slides

Capture of genomic DNA on glass microscope slides

Review on Impedance Detection of Cellular Responses in Micro/Nano Environment

Development of Capillary Loop Convective Polymerase Chain Reaction Platform with Real-Time Fluorescence Detection

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