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Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification
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Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification

Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification

DOI:
10.3791/62295-v

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07:54 min

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March 31, 2021

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, , , , , , , , , ,
1Key Laboratory of Special Pathogens and Biosafety, Center for Biosafety Mega-Science, Wuhan Institute of Virology,Chinese Academy of Sciences, 2International College,University of Chinese Academy of Sciences, 3Sino-Africa Joint Research Center, 4CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai,Chinese Academy of Sciences, 5Center for Infectious and Parasitic Diseases Control Research,Kenya Medical Research Institute

Chapters

  • 00:04Introduction
  • 00:48cDNA and Droplet Digital Polymerase Chain Reaction (ddPCR) Preparation
  • 02:03Automated Droplet Generation
  • 02:59PCR Amplification and Droplet Reading
  • 04:09Data Analysis
  • 06:01Results: SARS‐CoV‐2 Detection Using a Two‐Color ddPCR System
  • 07:07Conclusion

Summary

Automatic Translation

Automatic Translation

This work summarizes steps on developing different assays for SARS-CoV-2 detection using a two color ddPCR system. The steps are elaborate and notes have been included on how to improve the assays and experiment performance. These assays may be used for multiple SARS-CoV-2 RT-ddPCR applications.

Tags

Keyword Extraction:RT-ddPCRSARS-CoV-2Multiplex DdPCR AssayTwo-color DdPCR SystemRT-qPCRCDNA SynthesisReverse TranscriptionDroplet Digital PCRAutomated Droplet GeneratorMaster MixSample PlateDroplet Generation

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