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.

中文翻译：

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一种使用Taq DNA聚合酶的酶逆转录qPCR

Taq DNA聚合酶是最早发现的热稳定DNA聚合酶之一，已被定型为通常用于PCR的DNA依赖性DNA聚合酶。但是，Taq聚合酶与Molony鼠白血病病毒逆转录酶属于同一DNA聚合酶超家族，并且在过去已证明具有逆转录酶活性。我们报告了优化的缓冲液和盐成分，可促进Taq DNA聚合酶的逆转录酶活性，从而使其可用作TaqMan RT-qPCR反应中的唯一酶。我们通过执行CDC SARS-CoV-2 N1，N2和N3 TaqMan RT-qPCR分析来证明仅Taq的RT-qPCR反应的实用性，该检测可以检测到低至2拷贝/μL的输入病毒基因组RNA。