Electrical Detection of Nucleic Acid Amplification Using an On-Chip Quasi-Reference Electrode and a PVC REFET
| Autor: | Vikhram V. Swaminathan, Rashid Bashir, Carlos Duarte-Guevara, Bobby Reddy, Yu Zhong, Eric Salm, Yi Shao Liu |
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| Rok vydání: | 2014 |
| Předmět: |
Chemistry
Transistor Loop-mediated isothermal amplification Nanotechnology Equipment Design Hydrogen-Ion Concentration Polymerase Chain Reaction Reference electrode Article Analytical Chemistry law.invention Membrane law Electrode Nucleic acid Miniaturization Microtechnology Field-effect transistor Polyvinyl Chloride Electrodes Nucleic Acid Amplification Techniques Oligonucleotide Array Sequence Analysis Platinum |
| Zdroj: | Analytical Chemistry |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/ac500897t |
| Popis: | Electrical detection of nucleic acid amplification through pH changes associated with nucleotide addition enables miniaturization, greater portability of testing apparatus, and reduced costs. However, current ion-sensitive field effect transistor methods for sensing nucleic acid amplification rely on establishing the fluid gate potential with a bulky, difficult to microfabricate reference electrode that limits the potential for massively parallel reaction detection. Here we demonstrate a novel method of utilizing a microfabricated solid-state quasi-reference electrode (QRE) paired with a pH-insensitive reference field effect transistor (REFET) for detection of real-time pH changes. The end result is a 0.18 μm, silicon-on-insulator, foundry-fabricated sensor that utilizes a platinum QRE to establish a pH-sensitive fluid gate potential and a PVC membrane REFET to enable pH detection of loop mediated isothermal amplification (LAMP). This technique is highly amendable to commercial scale-up, reduces the packaging and fabrication requirements for ISFET pH detection, and enables massively parallel droplet interrogation for applications, such as monitoring reaction progression in digital PCR. |
| Databáze: | OpenAIRE |
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