Nearly Instantaneous, Cation-Independent, High Selectivity Nucleic Acid Hybridization to DNA Microarrays

Belosludtsev, Yuri; Belosludtsev, Inna; Iverson, Bonnie; Lemeshko, Sergy V.; Wiese, Rick; Hogan, Michael E.; Powdrill, Tom

doi:10.1006/bbrc.2001.4728

Cited by 25 publications

(13 citation statements)

References 7 publications

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“…The positively charged PLL layer seems to compensate the negative charge on the probe DNA and reduce the electrostatic repulsion between target and probe DNA. In addition, a positively charged surface may increase the local concentration of target oligonucleotides near the surface (29). The method of hybridizing nucleic acids on a charge-compensated surface at low ionic strength has been investigated recently for use in rapid microarray hybridization (29).…”

Section: Resultsmentioning

confidence: 99%

“…In addition, a positively charged surface may increase the local concentration of target oligonucleotides near the surface (29). The method of hybridizing nucleic acids on a charge-compensated surface at low ionic strength has been investigated recently for use in rapid microarray hybridization (29). It might also be used for other field-effect sensing devices such as silicon nanowires (30), which could lead to a dramatic size reduction of nucleic acid sensors.…”

Section: Resultsmentioning

confidence: 99%

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Electronic detection of DNA by its intrinsic molecular charge

Fritz

Cooper

Gaudet

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

428

320

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We report the selective and real-time detection of label-free DNA using an electronic readout. Microfabricated silicon field-effect sensors were used to directly monitor the increase in surface charge when DNA hybridizes on the sensor surface. The electrostatic immobilization of probe DNA on a positively charged poly-L-lysine layer allows hybridization at low ionic strength where field-effect sensing is most sensitive. Nanomolar DNA concentrations can be detected within minutes, and a single base mismatch within 12-mer oligonucleotides can be distinguished by using a differential detection technique with two sensors in parallel. The sensors were fabricated by standard silicon microtechnology and show promise for future electronic DNA arrays and rapid characterization of nucleic acid samples. This approach demonstrates the most direct and simple translation of genetic information to microelectronics. Awide range of techniques for detecting nucleic acids is based on their hybridization to DNA probes on a solid surface (ref. 1, entire issue, and ref. 2). In the methods used most routinely, the physical nature of the readout requires the attachment of reporter molecules such as fluorescent, chemiluminescent, redox, or radioactive labels (1, 3, 4). Although labeldependent methods achieve the highest sensitivities (5-7), eliminating the labeling steps has the advantage of simplifying the readout and increasing the speed and ease of nucleic acid assays, which is especially desirable for characterizing infectious agents, scoring sequence polymorphisms and genotypes, and measuring mRNA levels during expression profiling. The development of label-independent methods that can monitor hybridization in real time and that are simple and scalable is still in its infancy (8-11). Here we describe a label-free method for electronically detecting DNA by its intrinsic molecular charge using microfabricated field-effect sensors.The field-effect sensor is based on an electrolyte-insulator-silicon (EIS) structure. Variations in the insulator-electrolyte surface potential, which arise from the binding of charged molecules (e.g., nucleic acids) to the insulator surface ( Fig. 1 a and b), modify the charge distribution in the silicon below the electrolyte. Surface charge and surface potential at this interface are related according to the Grahame equation (12). The surface potential can be measured by changes in conductivity (13) or capacitance (14) in the silicon part of the EIS structure. We have chosen to measure the capacitance, because it requires only one electrical connection to the silicon. The measured capacitance between the silicon and counter electrode in the electrolyte solution is dominated by the insulator capacitance and the capacitance of the charge-depleted region in the silicon. These two capacitances appear in series, and only the silicon depletion capacitance is modulated by the insulator surface potential. The capacitance-versus-voltage dependence of EIS structures (15) is similar to that of metal-oxide-semiconducto...

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Electronic detection of DNA by its intrinsic molecular charge

Fritz

Cooper

Gaudet

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

428

320

View full text Add to dashboard Cite

show abstract

“…Different substrate chemistries are in focus and methods are being developed for efÞ cient binding of oligonucleotides to different kinds of substrate. [28,29] In our hands, the aldehyde reactive surface showed maximum binding capacity for amino linked oligonucleotides. In addition, use of dT (15) spacer overcomes the steric hindrance of the surface structure and solvent layer due to its charge, hydrophobicity, degree of solvation.…”

Section: Discussionmentioning

confidence: 60%

Fabrication and evaluation of a sequence-specific oligonucleotide miniarray for molecular genotyping

Iqbal

Hänel

Ruryk

et al. 2008

Indian J Med Microbiol

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“…For example, in DNA microarrays, where the capture probe has been functionalised at its terminus with biotin and anchored to a surface coated with streptavidin, at low ionic strengths and at pH 6, where streptavidin carries a positive charge, hybridisation of a target sequence at a 1 nanomolar concentration is at least 80-fold quicker than under neutral pH and higher ionic strength conditions [59].…”

Section: Miniaturisation and Scaling Laws: Nanoscale Devices And Perfmentioning

confidence: 99%

Biosensor Design with Molecular Engineering and Nanotechnology

Johnson

Trzebinski

et al. 2014

Body Sensor Networks

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The concept of a biosensor is well established and the idea of integrating a molecular recognition layer with a base sensor, such that analyte binding or reaction at the former results in a measureable change (in current or voltage) in the latter has seen many ingenious embodiments. Despite this and the huge amount of research worldwide in biosensors, as outlined in Chap. 2, many challenges still remain in building reliable, long-lived biosensors, especially in the hostile environment of the human body. The enormous potential for in vivo sensing of pathophysiological molecules over time and space has led to many attempts to achieve this and the rewards, both in terms of clinical benefit and improved understanding, cannot be underestimated. As new tools for producing biosensors become available, they are rapidly recruited. In recent years, developments in two areas of science and engineering have provided new opportunities to look again at how biosensors are built and deployed. These developments were not driven by the needs of those building and using biosensors but by much broader scientific and technological trends, which nonetheless have found ready applicability in this area.One of the trends is an increasing knowledge of the structural factors that determine function in biological macromolecules and the other is the appreciation that the properties of materials with a characteristic length scale from 1 to 100 nm are not those expected from dividing macroscopic materials into smaller pieces nor those expected from adding atoms or molecules together. The first of these trends is sometimes referred to as biomolecular engineering and the second as nanotechnology.There are many drivers for the use of molecular engineering and nanotechnology in the design and application of biosensors and the past decade has seen these tools

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Nearly Instantaneous, Cation-Independent, High Selectivity Nucleic Acid Hybridization to DNA Microarrays

Cited by 25 publications

References 7 publications

Electronic detection of DNA by its intrinsic molecular charge

Electronic detection of DNA by its intrinsic molecular charge

Fabrication and evaluation of a sequence-specific oligonucleotide miniarray for molecular genotyping

Biosensor Design with Molecular Engineering and Nanotechnology

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