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One-Enzyme Reverse Transcription qPCR Using Taq DNA Polymerase.

ABSTRACT: 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-qPCRs. We demonstrate the utility of Taq-alone RT-qPCRs by executing CDC SARS-CoV-2 N1, N2, and N3 TaqMan RT-qPCR assays that could detect as few as 2 copies/?L of input viral genomic RNA.

SUBMITTER: Bhadra S

PROVIDER: S-EPMC7757722 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature

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One-Enzyme Reverse Transcription qPCR Using Taq DNA Polymerase.

Bhadra Sanchita S Maranhao Andre C AC Paik Inyup I Ellington Andrew D AD

Biochemistry 20201204 49

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 all ...[more]

PMID: 33275410

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