| Source: |
Purified from E. coli containing a recombinant plasmid encoding the E. coli Mug protein. |
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| Unit Definition: |
One unit cleaves 1 pmole of a 32P-labeled oligonucleotide probe containing 3,N4-ethenocytosine within an oligonucleotide duplex in one hour at 37°C.
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| Assay Conditions: |
1X REC Buffer 6 (20 mM Tris-Cl (pH 8.0), 0.1 mg/ml BSA, 1 mM EDTA, and 1 mM DTT), 1 pmole 3,N4-ethenocytosine oligonucleotide (Cat# 3864-100-01) labeled with 32P, 1 pmole complement D (Cat# 3849-100-04), and serial dilutions of enzyme in a reaction volume of 20 µl, incubated for 1 hour at 37°C.
gel electrophoresis.
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| Specificity: |
E. coli Mug catalyzes the excision of 3,N4-ethenocytosine, a form of DNA damage, in double or single stranded DNA. It also acts to excise uracil in uracil-guanine mismatches.
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| Storage: |
Freeze in working aliquots at -20°C in a manual defrost freezer to avoid repeated freeze-thawings. The enzyme is supplied in 25 mM HEPES (pH 7.6), 0.5 mM EDTA, 1 mM DTT, 1 mM PMSF, and 10% (v/v) glycerol. |
