A modular microfluidic system for deoxyribonucleic acid identification by short tandem repeat analysis

Reedy, Carmen R.; Hagan, Kristin A.; Marchiarullo, Daniel J.; Dewald, Alison H.; Barron, Annelise E.; Bienvenue, Joan M.; Landers, James P.

doi:10.1016/j.aca.2010.12.016

Cited by 11 publications

(8 citation statements)

References 26 publications

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“…Following this, voltages ranging from 200 to −1800 V were applied to this domain in order to electrophoretically separate the DNA fragments. The device supported this process with no observed issues, and the DNA fragments were separated successfully, with the profile not looking dissimilar to that obtained in a glass device [ 14 ]. This result also demonstrated the compatibility of the HSA material with the gold leaf electrodes, which had thus far only been integrated into PeT/ink toner devices.…”

Section: Resultsmentioning

confidence: 95%

Rapid Fabrication of Electrophoretic Microfluidic Devices from Polyester, Adhesives and Gold Leaf

et al. 2017

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In the last decade, the microfluidic community has witnessed an evolution in fabrication methodologies that deviate from using conventional glass and polymer-based materials. A leading example within this group is the print, cut and laminate (PCL) approach, which entails the laser cutting of microfluidic architecture into ink toner-laden polyester sheets, followed by the lamination of these layers for device assembly. Recent success when applying this method to human genetic fingerprinting has highlighted that it is now ripe for the refinements necessary to render it amenable to mass-manufacture. In this communication, we detail those modifications by identifying and implementing a suitable heat-sensitive adhesive (HSA) material to equip the devices with the durability and resilience required for commercialization and fieldwork. Importantly, this augmentation is achieved without sacrificing any of the characteristics which make the PCL approach attractive for prototyping. Exemplary HSA-devices performed DNA extraction, amplification and separation which, when combined, constitute the complete sequence necessary for human profiling and other DNA-based analyses.

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Section: Resultsmentioning

confidence: 95%

Rapid Fabrication of Electrophoretic Microfluidic Devices from Polyester, Adhesives and Gold Leaf

et al. 2017

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“…The separation technique is widely used in different applications, and nowadays it is almost routinely used for DNA separation and quantification [56][57][58][59]. While sensitivity enhancements of more than 100 000-fold were reported with EKS in capillaries, lower but quite impressive figures were obtained when moving to microchip platform.…”

Section: Eks In Microchip Electrophoresismentioning

confidence: 97%

“…Driven by the increasing need for portable devices and very fast analysis times, microchip electrophoresis has gained massive publicity in the past two decades 54, 55. The separation technique is widely used in different applications, and nowadays it is almost routinely used for DNA separation and quantification 56–59. While sensitivity enhancements of more than 100 000‐fold were reported with EKS in capillaries, lower but quite impressive figures were obtained when moving to microchip platform.…”

Section: Stacking By Eksmentioning

confidence: 99%

High‐sensitivity capillary and microchip electrophoresis using electrokinetic supercharging

Dawod

Chung

2011

J of Separation Science

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Electrokinetic supercharging (EKS) is considered as one of the most powerful online preconcentration techniques in electrophoresis. It combines the efficient preconcentration power of field-amplified sample injection and the exceptional selective nature of transient isotachophoresis. It has a wide range of applications to different types of analytes ranging from small ions to large proteins and DNA fragments. This comprehensive review--up to date--provides listing for all the works, developments, and advances in EKS. The review will pay particular attention to innovations, new methodologies for manipulation, challenges for improving the detection sensitivity, and various applications of EKS in capillaries and microchips.

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“…Alternatively, microchip‐based μCE takes place in a fraction of the time in microcapillaries that are much shorter in length. Researchers have developed a μCE module and detection/separation system that, when combined with modules for SPE and IR‐PCR, successfully generates STR profiles equivalent to those run in parallel using conventional instruments . In the current work described herein, we did not attempt to integrate μCE onto the microdevice for two reasons.…”

Section: Introductionmentioning

confidence: 99%

A novel, integrated forensic microdevice on a rotation‐driven platform: Buccal swab to STR product in less than 2 h

et al. 2016

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This work describes the development of a novel microdevice for forensic DNA processing of reference swabs. This microdevice incorporates an enzyme‐based assay for DNA preparation, which allows for faster processing times and reduced sample handling. Infrared‐mediated PCR (IR‐PCR) is used for STR amplification using a custom reaction mixture, allowing for amplification of STR loci in 45 min while circumventing the limitations of traditional block thermocyclers. Uniquely positioned valves coupled with a simple rotational platform are used to exert fluidic control, eliminating the need for bulky external equipment. All microdevices were fabricated using laser ablation and thermal bonding of PMMA layers. Using this microdevice, the enzyme‐mediated DNA liberation module produced DNA yields similar to or higher than those produced using the traditional (tube‐based) protocol. Initial microdevice IR‐PCR experiments to test the amplification module and reaction (using Phusion Flash/SpeedSTAR) generated near‐full profiles that suffered from interlocus peak imbalance and poor adenylation (significant −A). However, subsequent attempts using KAPA 2G and Pfu Ultra polymerases generated full STR profiles with improved interlocus balance and the expected adenylated product. A fully integrated run designed to test microfluidic control successfully generated CE‐ready STR amplicons in less than 2 h (<1 h of hands‐on time). Using this approach, high‐quality STR profiles were developed that were consistent with those produced using conventional DNA purification and STR amplification methods. This method is a smaller, more elegant solution than current microdevice methods and offers a cheaper, hands‐free, closed‐system alternative to traditional forensic methods.

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A modular microfluidic system for deoxyribonucleic acid identification by short tandem repeat analysis

Cited by 11 publications

References 26 publications

Rapid Fabrication of Electrophoretic Microfluidic Devices from Polyester, Adhesives and Gold Leaf

Rapid Fabrication of Electrophoretic Microfluidic Devices from Polyester, Adhesives and Gold Leaf

High‐sensitivity capillary and microchip electrophoresis using electrokinetic supercharging

A novel, integrated forensic microdevice on a rotation‐driven platform: Buccal swab to STR product in less than 2 h

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