Plug-and-play, infrared, laser-mediated PCR in a microfluidic chip

Pak, Nikita; Saunders, D. Curtis; Phaneuf, Christopher R.; Forest, Craig R.

doi:10.1007/s10544-011-9619-2

Cited by 24 publications

(20 citation statements)

References 32 publications

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“…52 The reported power consumption for infrared systems is similar to Peltier systems, at 4-5 W per well. The infrared energy is differentially absorbed by the aqueous solution and the surrounding plastic tube or chip.…”

Section: View Article Onlinementioning

confidence: 78%

Portable nucleic acid thermocyclers

2013

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A nucleic acid thermal cycler is considered to be portable if it is under ten pounds, easily carried by one individual, and battery powered. Nucleic acid amplification includes both polymerase chain reaction (e.g. PCR, RT-PCR) and isothermal amplification (e.g. RPA, HDA, LAMP, NASBA, RCA, ICAN, SMART, SDA). There are valuable applications for portable nucleic acid thermocyclers in fields that include clinical diagnostics, biothreat detection, and veterinary testing. A system that is portable allows for the distributed detection of targets at the point of care and a reduction of the time from sample to answer. The designer of a portable nucleic acid thermocycler must carefully consider both thermal control and the detection of amplification. In addition to thermal control and detection, the designer may consider the integration of a sample preparation subsystem with the nucleic acid thermocycler. There are a variety of technologies that can achieve accurate thermal control and the detection of nucleic acid amplification. Important evaluation criteria for each technology include maturity, power requirements, cost, sensitivity, speed, and manufacturability. Ultimately the needs of a particular market will lead to user requirements that drive the decision between available technologies.

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Section: View Article Onlinementioning

confidence: 78%

Portable nucleic acid thermocyclers

2013

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“…As described in the previous work, 25,31,32 the microchips were fabricated from PMMA substrates using a 3-axis vertical milling center (Haas) at a rate of two chips per minute. The 60 mm Â 20 mm substrates were laser cut from sheets of 1.5 mm thick cast PMMA.…”

Section: Microfluidic Devicementioning

confidence: 99%

“…Techniques used in the previous work include reference chambers with thermocouples, 19 fluorescent dyes, 22 pyrometers, 24 resistance measurements, 15 and open-loop control. 25,26 Most PCR technologies, especially within the field of microfluidics, have advanced the speed and throughput for screening many samples or patients for a single genetic target. For multiple targets, a biochemical technique called multiplex PCR was developed in the 1980s (Ref.…”

Section: Introductionmentioning

confidence: 99%

Thermally multiplexed polymerase chain reaction

et al. 2015

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Amplification of multiple unique genetic targets using the polymerase chain reaction (PCR) is commonly required in molecular biology laboratories. Such reactions are typically performed either serially or by multiplex PCR. Serial reactions are time consuming, and multiplex PCR, while powerful and widely used, can be prone to amplification bias, PCR drift, and primer-primer interactions. We present a new thermocycling method, termed thermal multiplexing, in which a single heat source is uniformly distributed and selectively modulated for independent temperature control of an array of PCR reactions. Thermal multiplexing allows amplification of multiple targets simultaneously-each reaction segregated and performed at optimal conditions. We demonstrate the method using a microfluidic system consisting of an infrared laser thermocycler, a polymer microchip featuring 1 ll, oil-encapsulated reactions, and closed-loop pulsewidth modulation control. Heat transfer modeling is used to characterize thermal performance limitations of the system. We validate the model and perform two reactions simultaneously with widely varying annealing temperatures (48 C and 68 C), demonstrating excellent amplification. In addition, to demonstrate microfluidic infrared PCR using clinical specimens, we successfully amplified and detected both influenza A and B from human nasopharyngeal swabs. Thermal multiplexing is scalable and applicable to challenges such as pathogen detection where patients presenting non-specific symptoms need to be efficiently screened across a viral or bacterial panel. V C 2015 AIP Publishing LLC.

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“…Some studies have developed the chamber type PCR devices based on the conception of a conventional thermocycler. The mixture is kept static, and the temperature of the reaction chamber is cycled [2]. Some researchers have examined the continuous flow PCR (CFPCR) devices, and the reaction mixture moves through one or several different isothermal regions repetitively in a microchannel or tube [3].…”

Section: Introductionmentioning

confidence: 99%

Analysis of Thermal Performance in a Bidirectional Thermocycler by Including Thermal Contact Characteristics

Chen

et al. 2014

Micromachines

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This paper illustrates an application of a technique for predicting the thermal characteristics of a bidirectional thermocycling device for polymerase chain reaction (PCR). The micromilling chamber is oscillated by a servo motor and contacted with different isothermal heating blocks to successfully amplify the DNA templates. Because a comprehensive database of contact resistance factors does not exist, it causes researchers to not take thermal contact resistance into consideration at all. We are motivated to accurately determine the thermal characteristics of the reaction chamber with thermal contact effects existing between the heater surface and the chamber surface. Numerical results show that the thermal contact effects between the heating blocks and the reaction chamber dominate the temperature variations and the ramping rates inside the PCR chamber. However, the influences of various temperatures of the ambient conditions on the sample temperature during three PCR steps can be negligible. The experimental temperature profiles are compared well with the numerical simulations by considering the thermal contact conductance coefficient which is empirical by the experimental fitting. To take thermal contact conductance coefficients into consideration in the thermal simulation is recommended to predict a reasonable temperature profile of the reaction chamber during various thermal cycling processes. Finally, the PCR experiments present that Hygromycin B DNA templates are amplified successfully. Furthermore, our group is the first group to OPEN ACCESS Micromachines 2014, 5 1446 introduce the thermal contact effect into theoretical study that has been applied to the design of a PCR device, and to perform the PCR process in a bidirectional thermocycler.

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Plug-and-play, infrared, laser-mediated PCR in a microfluidic chip

Cited by 24 publications

References 32 publications

Portable nucleic acid thermocyclers

Portable nucleic acid thermocyclers

Thermally multiplexed polymerase chain reaction

Analysis of Thermal Performance in a Bidirectional Thermocycler by Including Thermal Contact Characteristics

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