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Can Uracil-N-Glycosylase (UNG) be used with a one-step RT-PCR kit, such as the QuantiTect Probe RT-PCR Kit?
No. Uracil-N-Glycosylase will remain active during the reverse-transcription step with the QuantiTect Probe RT-PCR Kit, and destroy any newly synthesized cDNA. Heat-labile UNG is required for one-step RT-PCR. See also FAQ 564 for further info.
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QuantiTect RT-PCR Kits
QuantiTect RT-PCR Kits enable sensitive quantification of RNA targets by real-time one-step PCR using sequence-specific probes or SYBR Green I detection. The kits also allow reliable quantification of up to 5 RNA targets in a single tube by multiplex, real-time one-step RT-PCR. The combination of a hot start and a unique PCR buffer system in the ready-to-use master mix ensures highly sensitive qRT-PCR on any real-time cycler without the need for optimization. The dNTP mix includes dUTP, allowing optional treatment with UNG. For convenience, the master mixes in QuantiTect one-step RT-PCR kits can be stored at 2–8°C. Two kit formats are available for multiplex RT-PCR using sequence-specific probes: the QuantiTect Multiplex RT-PCR Kit for cyclers that require ROX dye for fluorescence normalization, and the QuantiTect Multiplex RT-PCR NoROX Kit for all other cyclers. The QuantiTect SYBR Green RT-PCR Kit is supplied with an optimized RT mix for efficient and sensitive reverse transcription over a wide range of RNA template amounts.