Uracil-DNA Excision Mix

Applications

  • Mapping of uracil-containing residues in any DNA.
  • Mapping CpG islands.1-3
  • DNA repair studies.

Epicentre's Uracil-DNA Excision Mix is a blend of enzymes that cleave DNA at positions where uracil is present in place of thymine. The Uracil-DNA Excision Mix is useful for specific or random cleavage of DNA or for DNA repair studies, allowing mapping of uracil residues in any DNA. Uracil-DNA glycosylase in the Excision Mix removes uracil bases from DNA, creating a single-base gap and leaving the deoxyribose phosphate backbone intact. Endonuclease IV in the Excision Mix then cleaves the DNA at each abasic site, leaving a 3´-hydroxyl end and an abasic 5´-phosphorylated end. The minimum oligomer size that will serve as a substrate is six base pairs. Uracil-DNA Excision Mix digestion products can be analyzed by denaturing agarose gel electrophoresis or denaturing polyacrylamide gel electrophoresis.

Uracil Excision Enzyme 10X Buffer: 500 mM Tris-HCl (pH 9.0), 200 mM (NH4)2SO4, and 100 mM EDTA.

Quality Control: Uracil Excision Enzyme Mix activity is tested for activity on a variety of DNA templates, and is free of detectable RNase and nonspecific DNase activities.

References

  1. Peden, K.W. and Nathans, D. (1982) Proc. Natl. Acad. Sci. USA 79, 7214.
  2. Kubareva, E.A. et al. (2002) Biotechniques 33, 526.
  3. Morgan, A.R. et al. (1988) Biochem. Cell Biol. 66, 157.