| Source: |
Purified from E. coli containing a recombinant plasmid harboring the E. coli nei gene. |
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| Unit Definition: |
One unit is the amount of enzyme required to cleave an AP site oligonucleotide within an oligonucleotide duplex at the rate of 1 pmol/hour at 37°C.
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| Assay Conditions: |
1X REC Buffer 3 (10 mM HEPES-KOH (pH 7.4), 100 mM KCl, and 1 mM EDTA), 1 pmole of an AP-site Oligonucleotide labeled with 32P, 1 pmole of complementary oligonucleotide, and serial dilutions of enzyme in a 20 µl reaction volume are incubated for 1 hour at 37°C. Cleavage products are resolved by 20% denaturing polyacrylamide gel electrophoresis. The bands are cut out and radioactivity is counted to quantify the cleavage products.
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| Applications: |
CometAssay®/FLARETM
Supercoiled DNA relaxation assay
Alkaline elution assay
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| Storage: |
Freeze in working aliquots at -20°C in a manual defrost freezer to avoid repeated freeze-thaws. The enzyme is supplied in buffer containing 50% glycerol. |
