Preconcentration and separation of double‐stranded DNA fragments by electrophoresis in plastic microfluidic devices

Wainright, Ann; Nguyen, Uyen; Björnson, Torleif O.; Boone, Travis

doi:10.1002/elps.200305594

Cited by 89 publications

(57 citation statements)

References 36 publications

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“…However, it is too specific for DNA molecules with a certain length due to its fixed pore size. Chemical method, 10,11 based on absorption and desorption processes, is another conventional way to enhance DNA concentration, prior to capillary electrophoresis (EP). Electrical method is the most popular technique due to its simple implementation and reliability.…”

Section: Introductionmentioning

confidence: 99%

On-chip DNA preconcentration in different media conductivities by electrodeless dielectrophoresis

Hui

et al. 2015

Biomicrofluidics

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Electrodeless dielectrophoresis is the best choice to achieve preconcentration of nanoparticles and biomolecules due to its simple, robust, and easy implementation. We designed a simple chip with microchannels and nano-slits in between and then studied the trapping of DNA in high conductive medium and low conductive medium, corresponding to positive and negative dielectrophoresis (DEP), respectively. It is very important to investigate the trapping in media with different conductivities since one always has to deal with the sample solutions with different conductivities. The trapping process was analyzed by the fluorescent intensity changes. The results showed that DNA could be trapped at the nano-slit in both high and low conductive media in a lower electric field strength (10 V/cm) compared to the existing methods. This is a significant improvement to suppress the Joule heating effect in DEP related experiments. Our work may give insight to researchers for DNA trapping by a simple and low cost device in the Lab-on-a-Chip system. V C 2015 AIP Publishing LLC. [http://dx

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Section: Introductionmentioning

confidence: 99%

On-chip DNA preconcentration in different media conductivities by electrodeless dielectrophoresis

Hui

et al. 2015

Biomicrofluidics

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“…The creation and integration of these components represents a more substantial problem than it might seem, but the development of new microfluidic elements, and means to incorporate them into devices, are proceeding. Systems for bioanalysis and separation, [1][2][3] or high-throughput screening 4,5 are already available. We are especially interested in devices to be used in resource poor environments-e.g., in healthcare in developing countries, by first responders and the military, and in analogous problems-where portability and ruggedness are key concerns.…”

Section: Introductionmentioning

confidence: 99%

Mixing with bubbles: a practical technology for use with portable microfluidic devices

et al. 2006

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This paper demonstrates a methodology for micromixing that is sufficiently simple that it can be used in portable microfluidic devices. It illustrates the use of the micromixer by incorporating it into an elementary, portable microfluidic system that includes sample introduction, sample filtration, and valving. This system has the following characteristics: (i) it is powered with a single hand-operated source of vacuum, (ii) it allows samples to be loaded easily by depositing them into prefabricated wells, (iii) the samples are filtered in situ to prevent clogging of the microchannels, (iv) the structure of the channels ensure mixing of the laminar streams by interaction with bubbles of gas introduced into the channels, (v) the device is prepared in a single-step soft-lithographic process, and (vi) the device can be prepared to be resistant to the adsorption of proteins, and can be used with or without surface-active agents.We fabricated the device (Figs. 1a) in a polydimethylsiloxane (PDMS) slab sealed to a PDMS substrate. We chose to use PDMS because it is the most useful material for testing concepts in microfluidics, and because it is easily coupled with

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“…A drawback of these techniques is that they allow only relatively small sample volumes (a few microliters) to be used, and the target molecules are not always nucleic acids. Transient ITP offers the advantage of size separation after preconcentration (34 ), which facilitates the concentration of a DNA sample before separation (35 ); however, transient ITP suffers from the drawback of requiring long channels that exceed the chip dimensions for samples Ͼ10 L. Other approaches for preconcentration have exploited filter structures or membranes (36,37 ) and, recently, electrokinetic trapping (EKT) at polyethylene terephthalate (PET) membranes for stacking larger volumes of about 80 L (38 ).…”

confidence: 99%

Microsystem for Isolation of Fetal DNA from Maternal Plasma by Preparative Size Separation

2009

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Background: Routine prenatal diagnosis of chromosomal anomalies is based on invasive procedures, which carry a risk of approximately 1%–2% for loss of pregnancy. An alternative to these inherently invasive techniques is to isolate fetal DNA circulating in the pregnant mother’s plasma. Free fetal DNA circulates in maternal plasma primarily as fragments of lengths <500 bp, with a majority being <300 bp. Separating these fragments by size facilitates an increase in the ratio of fetal to maternal DNA. Methods: We describe our development of a microsystem for the enrichment and isolation of cell-free fetal DNA from maternal plasma. The first step involves a high-volume extraction from large samples of maternal plasma. The resulting 80-μL eluate is introduced into a polymeric microsystem within which DNA is trapped and preconcentrated. This step is followed by a transient isotachophoresis step in which the sample stacks within a neighboring channel for subsequent size separation and is recovered via an outlet at the end of the channel. Results: Recovered fractions of fetal DNA were concentrated 4–8 times over those in preconcentration samples. With plasma samples from pregnant women, we detected the fetal SRY gene (sex determining region Y) exclusively in the fragment fraction of <500 bp, whereas a LEP gene (leptin) fragment was detected in both the shorter and longer recovery fractions. Conclusions: The microdevice we have described has the potential to open new perspectives in noninvasive prenatal diagnosis by facilitating the isolation of fetal DNA from maternal plasma in an integrated, inexpensive, and easy-to-use microsystem.

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Preconcentration and separation of double‐stranded DNA fragments by electrophoresis in plastic microfluidic devices

Cited by 89 publications

References 36 publications

On-chip DNA preconcentration in different media conductivities by electrodeless dielectrophoresis

On-chip DNA preconcentration in different media conductivities by electrodeless dielectrophoresis

Mixing with bubbles: a practical technology for use with portable microfluidic devices

Microsystem for Isolation of Fetal DNA from Maternal Plasma by Preparative Size Separation

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