| Autor: |
Elizabeth A. Phillips, Adam D. Silverman, Aric Joneja, Michael Liu, Carl Brown, Paul Carlson, Christine Coticchia, Kristen Shytle, Alex Larsen, Nadish Goyal, Vincent Cai, Jason Huang, Jennifer E. Hickey, Emily Ryan, Joycelynn Acheampong, Pradeep Ramesh, James J. Collins, William J. Blake |
| Rok vydání: |
2023 |
| DOI: |
10.21203/rs.3.pex-2197/v1 |
| Popis: |
This protocol defines the process for executing an INSPECTR assay to sensitively detect SARS-CoV-2 gRNA in a fully ambient temperature workflow. INSPECTR (for internal splint-pairing expression-cassette translation reaction), leverages the target-specific splinted ligation of DNA probes to generate expression cassettes that can be flexibly designed for the cell-free synthesis of reporter proteins. One implementation of INSPECTR incorporates rolling circle amplification, which is described in this protocol. This protocol entails five ambient temperature steps including target-splinted probe ligation, exonuclease digestion, hyperbranched rolling circle amplification, in vitro transcription and translation, and lateral flow detection. This implementation of the INSPECTR assay demonstrates the utility of synthetic biology and cell-free systems wherever low-cost and decentralized nucleic acid sensing is of value. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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