General description:

Uracil-DNA glycosylase (UDG) removes uracil from dU-containing DNA by cleaving the N-glycosidic bond between the uracil base and the sugar backbone. This cleavage generates an alkali-sensitive pyrimidine site that is blocked from replication by DNA polymerase or prevented from becoming a hybridization site. Only double-stranded and single-stranded dU-containing DNA can be recognized and degraded by uracil-DNA glycosylase, while RNA and normal dT-containing DNA are not recognized at all. Uracil-DNA glycosylase from cold-water fish is heat-labile. Complete irreversible inactivation occurs after incubation at 50°C for 10 minutes.

Definition of activity unit

● The amount of enzyme required to degrade 1 μg of dsDNA containing uracil within 30 minutes at 25°C is defined as 1 activity unit (U).

Applications:

● DNA repair and mutation detection.
● Improve the cloning efficiency of PCR products.
● Improve the efficiency of site-directed mutagenesis.
● Study of protein-DNA interactions.
● Eliminate aerosol contamination during PCR, qPCR and RT-qPCR amplification

Features and Benefits

● Prevents PCR carryover contamination.
● Improves efficiency during site-directed mutagenesis.
● Labels oligonucleotide probes.
● Faster inactivation than corresponding enzymes from E. coli due to lower thermal stability.
● Does not degrade dU-PCR products for at least several hours when incubated at +2 to +8°C.