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- W3131082807 abstract "In single molecule, real-time (SMRT) sequencing technology, individual nucleotides are monitored in real time during DNA strand replication by an individual DNA polymerase tracking the incorporation of multi-color fluorescently labeled nucleotides. The DNA template to be sequenced is loaded to the bottom of nanowells called zero-mode waveguides (ZMWs) via a random diffusion process, thereby minimizing throughput and read length. Also, fundamental size constraints of ZMWs hinder the loading of large molecules into ZMWs creating a length bias. In order to overcome these challenges, we recently developed electrically actuatable ZMWs, nanopore ZMWs (NZMWs) and porous ZMWs (PZMW), to electrokinetically enhance the loading of biomolecules at sub-ng levels. The efficiency of voltage-induced DNA loading into these waveguides was length-independent and is 6-7 orders of magnitude larger than diffusion-based SMRT sequencing. Although PZMWs offer a higher DNA capture rate compared to NZMWs, the longevity of the nanoporous inorganic layer is limited due to the erosion of the porous aluminum oxide upon reaction with the ambient atmosphere. One drawback of both NZMWs and PZMWs is the reliance on ultrathin free-standing membranes, which reduces the overall scalability for high-throughput sequencing applications. In this work, we have developed a novel design of ZMWs, called electro-optical ZMWs (eZMWs) that can be electrically excited to draw DNA/RNA molecules. In this device, an electric field is created by the application of a voltage through an embedded metallic electrode under the aluminum waveguides made directly on a fused silica substrate. Our new device has improved DNA/RNA loading efficiency as compared to ordinary ZMWs, enables scale-up fabrication, enhances stability by eliminating the need for freestanding membranes, and features reduced background optical noise. Future applications of this device will include direct RNA sequencing." @default.
- W3131082807 created "2021-03-01" @default.
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- W3131082807 date "2021-02-01" @default.
- W3131082807 modified "2023-10-16" @default.
- W3131082807 title "DNA/RNA SMRT Sequencing using Electro-Optical Waveguides" @default.
- W3131082807 doi "https://doi.org/10.1016/j.bpj.2020.11.1277" @default.
- W3131082807 hasPublicationYear "2021" @default.
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