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Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Biochemistry 2020 ; 59 (49): 4638-4645 Nephropedia Template TP
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One-Enzyme Reverse Transcription qPCR Using Taq DNA Polymerase #MMPMID33275410
Bhadra S; Maranhao AC; Paik I; Ellington AD
Biochemistry 2020[Dec]; 59 (49): 4638-4645 PMID33275410show ga
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/muL of input viral genomic RNA.
|Animals[MESH]
|COVID-19/*diagnosis/genetics/virology[MESH]
|Humans[MESH]
|Mice[MESH]
|Moloney murine leukemia virus/enzymology[MESH]
|RNA-Directed DNA Polymerase/*chemistry/genetics[MESH]