OverView
Cod Uracil-DNA Glycosylase (Cod UNG) from Atlantic Cod is the only commercially available UNG enzyme that is completely and irreversibly inactivated by moderate heat treatment. The enzyme is produced in a recombinant E. coli (ung-) strain that contains a modified Cod UNG gene.
Key Features:
- Heat-labile – Completely and irreversibly inactivated at 55°C
- Contamination control – ideal in applications below
- Use of Cod UNG makes contamination control possible in RT-PCR
- Does not degrade PCR product post-PCR. This makes downstream use of the PCR product possible
- High purity enzyme, tested free of contaminating nucleases
- Detergent free
- Post-PCR Analysis – Enables post-PCR analysis
Applications
Ideal for contamination control in
- PCR carry-over prevention
- RT-PCR
- RT-qPCR
- qPCR
- kPCR
- RT-LAMP
Heat-labile Uracil-DNA Glycosylase
There are several commercially available Uracil-DNA glycosylases on the market today. Most of them are of bacterial origin and work well if you have no intention to further analyze the PCR products post-PCR. However, if you want to store your PCR products for downstream analysis such as cloning and sequencing, the reactivation of UNG and subsequent degradation of your PCR products are a problem with most of the commercially available UNGs. Cod UNG from ArcticZymes is completely and irreversibly inactivated by heat thus ensuring that sample integrity is maintained long-term regardless of storage conditions.
This is illustrated in figure 1, below
Figures
Properties
Recommended Protocols
1. Contamination control in PCR, qPCR and one-step RT-qPCR
Cod UNG works in all commercially available master mixes.
Be sure that you have used dUTP containing dNTP mixes in your previous PCR experiments.
- Add 0.2 U Cod UNG directly to your 20 µl PCR reaction.
- pre-incubate for 5 min at room temperature.
- For RT-qPCR, reverse transcribe your RNA at 50-55°C.
- Run your PCR.
- Store your PCR product at -20°C or 4°C degrees.
2. Contamination control in RT-LAMP
Cod UNG is ideal for contamination control in RT-LAMP.
One unit of Cod UNG per 30 μl reaction is sufficient for removing even high concentrations of carry-over contamination.
- Ensure that you use dNTP mixes containing dUTP in your experiments.
- Check that the RT-LAMP reaction is compatible with dUTP by running side-by-side reactions containing different ratios of dUTP to dUTP (100% dUTP, 90% dUTP, 80% dUTP and 0% dUTP).
- Add 1 U Cod UNG directly to your 30 µl RT-LAMP reaction.
- Prepare the reaction mix on ice.
- Analyze your RNA at 65°C, no preincubation is necessary.
Please refer to Protocol for Carry-over Contamination Control
Ordering Info
