A transmission imaging spectrograph and microfabricated channel system for DNA analysis

Simpson, John; Ruiz‐Martinez, Marie C.; Mulhern, Greg T.; Berka, Jan; Latimer, Darin R.; D., Ball, James A.; Rothberg, Jonathan M.; Went, Greg T.

doi:10.1002/(sici)1522-2683(20000101)21:1<135::aid-elps135>3.0.co;2-u

Cited by 43 publications

(13 citation statements)

References 40 publications

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“…There are indications that this new instrumentation might soon become commercially available. In one approach, Simpson et al [84] at CuraGen Corporation (New Haven, CT, USA) have described a new microfabricated channel device containing 48 channels etched on a glass substrate. The bottom plate was made using Borofloat glass, into which 48 channels were etched to a depth of 50 mm.…”

Section: Microfabricated Capillary Arrays (ìCae)mentioning

confidence: 99%

Recent progress in DNA analysis by capillary electrophoresis

Righetti¹,

Gelfi²,

D’Acunto³

2002

Electrophoresis

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A number of recent developments in DNA analysis by capillary electrophoresis are here reviewed. They include capillary arrays for fast, parallel DNA sequencing as well as microfabricated capillary arrays. Microfluidic chips for DNA sizing and quantitation are also covered, as well as microdevices containing arrays of regular obstacles acting as size-separators during DNA migration. Screening of DNA point mutations by two much improved techniques is also reported: in one case, such mutations are detected (but only on relative short, ca. 60-70 base-long fragments) by free electrophoresis in rather acidic (pH ca. 3) buffers; in the case of single-strand chain polymorphism, an improved technique is described based on near-neutral pH buffers with mixtures of Tris/MES cations/zwitterions. When studying the behavior of inorganic and organic cations in the Debye-Hückel layer of DNA, it was found that the latter (especially a large number of Good's buffers and other zwitterions, such as His) would bind to the DNA filament not only via charge interaction, but also via additional bonds, notably hydrogen bonds, thus altering the electrophoretic (and possibly the biological) behavior of DNA molecules. However, whether or not borate ions would bind to DNA remains still an unsettled question. Finally, capillary electrophoresis was found to be instrumental in measuring fine physicochemical parameters pertaining to DNA polyelectrolytes, such as their free mobility and their translational diffusion coefficients.

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Section: Microfabricated Capillary Arrays (ìCae)mentioning

confidence: 99%

Recent progress in DNA analysis by capillary electrophoresis

Righetti¹,

Gelfi²,

D’Acunto³

2002

Electrophoresis

View full text Add to dashboard Cite

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“…The basic design of the separation system is similar to Simpson's design [44]. Briefly, a high-voltage power supply from Bertan (Hicksville, NY, USA) was used to drive electrophoresis.…”

Section: Equipmentmentioning

confidence: 99%

DNA analysis on microfabricated electrophoretic devices with bubble cells

et al. 2002

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Microfluidic devices with bubble cells have been fabricated on poly(methyl methacrylate) (PMMA) plates and have been employed for the analysis of DNA using polyethylene oxide (PEO) solutions. First, the separation channel was fabricated using a wire-imprinting method. Then, wires with greater sizes or a razor blade glued in a polycarbonate plate was used to fabricate bubble cells, with sizes of 190-650 microm. The improvements in resolution and sensitivity have been achieved for large DNA (> 603 base pair, bp) using such devices, which depend on the geometry of the bubble cell. The main contributor for optimal resolution is mainly due to DNA migration at lower electric field strengths inside the bubble cell. On the other hand, slight losses of resolution for small DNA fragments have been found mainly due to diffusion, supported by the loss of resolution when separating two small solutes. With a bubble cell of 75 microm (width) x 500 microm (depth), the sensitivity improvement up to 17-fold has been achieved for the 271 bp fragment in the separation of PhiX-174/HaeIII DNA restriction fragments. We have also found that a microfluidic device with a bubble cell of 360 microm x 360 microm is appropriate for DNA analysis. Such a device has been used for separating DNA ranging from 8 to 2176 bp and polymerase chain reaction (PCR) products amplified after 30 cycles, with rapidity and improvements in the sensitivity as well as resolution.

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“…Latest developments in microchip electrophoresis have been summarized in recent reviews [8][9][10]. One of the most promising features of microchip electrophoresis (MCE) is its great potential for high-throughput analysis due to the possibility for high-speed separations [11] and the ease of multiplexing the separation channels [12][13][14][15]. We have recently shown that capillary array electrophoresis and MCE have great potential as high-throughput screening (HTS) systems for assaying the enantioselectivity of large libraries of potential catalysts [16].…”

Section: Introductionmentioning

confidence: 99%

Design and performance of a microchip electrophoresis instrument with sensitive variable-wavelength fluorescence detection

et al. 2002

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A modular instrument for high-speed microchip electrophoresis (MCE) equipped with a sensitive variable-wavelength fluorescence detection system was developed and evaluated. The experimental setup consists mainly of a lamp-based epifluorescence microscope for variable-wavelength fluorescence detection and imaging and a programmable four-channel bipolar high-voltage source capable of delivering up to +/- 10 kV per channel. The optical unit was equipped with a high-sensitivity photomultiplier tube and an adjustable aperture. The system was applied to MCE separations of flurescein isothiocyanate (FITC)-labelled amines utilizing blue light (450-480 nm) for excitation as well as for the separation of rhodamines utilizing excitation light in the green spectral region (531-560 nm). At optimized conditions baseline separation of four FITC-labelled amines could be obtained in less than 50 s at a detection limit of 460 ppt (1 nM) with a signal-to-noise ratio of 3:1. Three rhodamines could be baseline-separated in less than 6 s at a detection limit of 240 ppt (500 pM). The relative standard deviations of absolute migration times determined in repetitive MCE separations of FITC-labelled amines were below 2.5% (n= 25). By the application of cyclodextrin-modified electrolytes, chiral separation of FITC-labelled amines could be performed in seconds demonstrating the potential of microchip electrophoresis for chiral high-throughput screening.

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A transmission imaging spectrograph and microfabricated channel system for DNA analysis

Cited by 43 publications

References 40 publications

Recent progress in DNA analysis by capillary electrophoresis

Recent progress in DNA analysis by capillary electrophoresis

DNA analysis on microfabricated electrophoretic devices with bubble cells

Design and performance of a microchip electrophoresis instrument with sensitive variable-wavelength fluorescence detection

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