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Merck
CN

KEM0011

Sigma-Aldrich

10X Uracil Cleavage System

Ultra-pure enzyme for nucleic acid modifications

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grade

Molecular Biology

Assay

>99% (SDS-PAGE)

form

buffered aqueous solution

shipped in

dry ice

storage temp.

−20°C

General description

The Uracil Cleavage System provides two enzymes, which, when added sequentially to a reaction containing a synthetic DNA fragment containing a deoxy-uracil, generate a single nucleotide gap at the location of the uracil residue. The system consists of two individual enzyme components, Uracil DNA Glycosylase (UDG) and Endonuclease VIII, provided at a 10X concentration to be added to a reaction containing a uracil-containing polynucleotide sequence. UDG catalyzes the excision of the uracil base, creating an abasic site with an intact phosphodiester backbone. The lyase activity of Endonuclease VII breaks the phosphodiester backbone both 3′ and 5′ to the abasic site, liberating the deoxyribose sugar.

Application

Suitable for excision of uracil residues and successive generation of a single nucleotide gap.

Features and Benefits

  • Ultra-purification process for ultimate enzyme performance
  • Highest quality specifications for ultimate product consistency
  • Undetectable DNA and nuclease contamination

Other Notes

Source of protein: Each component protein is purified separately from E. coli strains containing recombinant Endonuclease VIII and Uracil-DNA Glycosylase genes.
Supplied with:KEM0001B (UDG)KEM0007B (Endonuclease VIII)
UDG: 1 unit is defined as the amount of enzyme that catalyzes the release of 1.8 nmol of Uracil in 30 minutes from double-stranded, tritiated, Uracil containing-DNA at 37° C in 1X UDG Reaction Buffer. Endonuclease VII: 1 unit is defined as the amount of enzyme required to cleave 1 pmol of an oligonucleotide duplex containing a single AP site in 1 hour at 37° C.
Unit size: 750 reactions

Storage Class Code

10 - Combustible liquids

Regulatory Information

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Laure Rittié et al.
Journal of cell communication and signaling, 2(1-2), 25-45 (2008-09-04)
Since molecular cloning has become routine laboratory technique, manufacturers offer countless sources of enzymes to generate and manipulate nucleic acids. Thus, selecting the appropriate enzyme for a specific task may seem difficult to the novice. This review aims at providing

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