Description
Uracil-DNA Glycosylase cleaves uracil from dU-containing DNA by breaking the N-glycosidic bond between the uracil base and the sugar backbone. This cleavage generates alkali-sensitive apyrimidinic sites that DNA polymerase blocks from replication or prevents from becoming a hybridization site. Single-stranded and double-stranded dU-containing DNA are substrates for uracil-DNA glycosylase, whereas normal dT-containing RNA and DNA are not.
Uracil-DNA glycosylase can also be used to increase the cloning efficiency of PCR products that have incorporated dU-containing primers, as well as to increase the efficiency of site-directed mutagenesis. Uracil-DNA Glycosylase derived from Gadus morhua has all the attributes of the enzyme derived from E. coli with the added benefit of being heat-labile. It is completely and irreversibly inactivated after 10 minutes of incubation at 50°C.
Specifications
- Quantity: 100U
- Kind of product: Uracil DNA Glycosylase
- Enzyme: glycosylase
- Compatible buffer: storage buffer
Content and storage
Shipped on dry ice. Store at -20°C.
Purity:
The enzyme is chromatographically purified and analyzed for contaminating endonucleases and exonucleases.
Storage buffer:
50 mM Tris-HCl (pH 7.5), 100 mM NaCl, 0.5 mM EDTA, 1 mM DTT, 0.1% Triton X-100, 50% glycerol.
Test conditions:
The reaction mixture contains 70 mM Tris-HCl, pH 8.0, 10 mM NaCl, 1 mM EDTA, 100 µg/mL BSA, 3 H-dUTP-labeled DNA, and uracil-DNA glycosylase. Incubation is at 37°C for 1 hour.
Unit definition:
One unit is the amount of enzyme required to release 1 nmol of uracil from dU-containing DNA in one hour at 37°C.
Concentration: 1 unit/µL
Applications:
- Study of DNA repair and mutation detection.
- Increase the cloning efficiency of PCR products.
- Increase the efficiency of site-directed mutagenesis.
- Study of protein-DNA interactions.
Fountain:
Recombinant Gadus morhua (cod liver)
