Uracil-DNA Glycosylase (UDG)

Application: UDG is used for carry-over protection in PCR, qPCR, RT, RT-qPCR and Isothermal Assays like LAMP by removing uracil from single- or double-stranded DNA.

Please use 1 unit of UDG in a reaction volume of 50 µl. The final concentration of dUTP can be adjusted between 0.2 – 0.6 mmol (0.2 mmol dTTP can be added if desired).

Use 25 °C for 5 – 10 min for dU-containing template degradation, followed by 95 °C for 5 – 10 min for UDG inactivation before continuing with the amplification protocol. UDG inactivation can be combined with an initial template denaturation step.

Unit definition: One unit of activity is defined as the amount of enzyme required to catalyze the hydrolysis of 1 µg dU-containing dsDNA in 30 min at 25 °C.

Heat inactivation: UDG is inactivated at 95 °C for 5 – 10 min. For reactions that require a lower temperature, please use a heat-labile UDG variant.

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