One-Enzyme Reverse Transcription qPCR Using Taq DNA Polymerase
| Autor: | Andre C. Maranhao, Inyup Paik, Andrew D. Ellington, Sanchita Bhadra |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
DNA polymerase
Biochemistry Article 03 medical and health sciences chemistry.chemical_compound Mice Murine leukemia virus TaqMan Animals Humans Taq Polymerase chemistry.chemical_classification 0303 health sciences biology Reverse Transcriptase Polymerase Chain Reaction SARS-CoV-2 RNA-Directed DNA Polymerase 030302 biochemistry & molecular biology COVID-19 biology.organism_classification Molecular biology Reverse transcriptase Enzyme chemistry biology.protein Moloney murine leukemia virus Taq polymerase Taq DNA Polymerase |
| Zdroj: | Biochemistry |
| ISSN: | 1520-4995 |
| Popis: | Taq DNA polymerase, one of the first thermostable DNA polymerases to be discovered, has been typecast as a DNA-dependent DNA polymerase commonly employed for PCR. However, Taq polymerase belongs to the same DNA polymerase superfamily as the Molony murine leukemia virus reverse transcriptase and has in the past been shown to possess reverse transcriptase activity. We report optimized buffer and salt compositions that promote the reverse transcriptase activity of Taq DNA polymerase, and thereby allow it to be used as the sole enzyme in TaqMan RT-qPCR reactions. We demonstrate the utility of Taq-alone RT-qPCR reactions by executing CDC SARS-CoV-2 N1, N2, and N3 TaqMan RT-qPCR assays that could detect as few as 2 copies/μL of input viral genomic RNA. |
| Databáze: | OpenAIRE |
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