Highly Efficient Real-Time Droplet Analysis Platform for High-Throughput Interrogation of DNA Sequences by Melt
| Autor: | Aniruddha M. Kaushik, Tza-Huei Wang, Christine O’Keefe |
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| Rok vydání: | 2019 |
| Předmět: |
Time Factors
Surface Properties Drug discovery Chemistry 010401 analytical chemistry Microfluidics Cysteine Dioxygenase High-Throughput Nucleotide Sequencing Large range Bacterial persistence Microfluidic Analytical Techniques 010402 general chemistry Polymerase Chain Reaction 01 natural sciences DNA sequencing 0104 chemical sciences Analytical Chemistry Antibody production Humans Transition Temperature Digital polymerase chain reaction Particle Size Biological system Throughput (business) |
| Zdroj: | Analytical Chemistry. 91:11275-11282 |
| ISSN: | 1520-6882 0003-2700 |
| Popis: | Droplet microfluidic platforms have greatly enhanced the throughput and sensitivity of single-molecule and single-cell analyses. However, real-time analyses of individual droplets remain challenging. Most droplet microfluidic platforms have fundamental drawbacks that undermine their utility toward applications that rely on real-time monitoring to identify rare variants, such as bacterial persistence, drug discovery, antibody production, epigenetic biomarker analyses, etc. We present a platform for high-density droplet trapping and real-time analysis with 100% loading and trapping efficiency at a packing density of 110,000 droplets per in2. To demonstrate real-time analysis capabilities, we perform digital PCR and parallelized digital high-resolution melt curve acquisition on droplets to discriminate methylation levels of a tumor suppressor gene, CDO1, on a molecule-by-molecule basis. We hope that this platform, which is compatible with a large range of droplet sizes and generation technologies, may facilitate high-throughput real-time analyses on a molecule-by-molecule or cell-by-cell basis of heterogeneous populations. |
| Databáze: | OpenAIRE |
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