| Source: |
Thermostable TDG is purified from E. coli containing a recombinant plasmid harboring the thermostable TDG gene. |
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| Unit Definition: |
One unit is the amount of enzyme required to cleave 1 pmole of an oligonucleotide duplex containing a T/G mismatch in 1 hour at 65°C. Only the strand containing the T is cleaved.
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| Assay Conditions: |
1X REC Buffer 4 (10 mM HEPES-KOH (pH 7.4), 100 mM KCl, and 10 mM EDTA), 4 pmoles of each oligo in the T/A Control Thermophilic Oligonucleotide Set (Cat# 3810-100-TA: TUT=TDG probe, TLA=complementary strand, and TLG=negative control), and serial dilutions of enzyme in a 20 µl reaction volume are incubated for 1 hour at 65°C.
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| Specificity: |
TDG enzyme recognizes T/G, mismatches in duplex DNA and cleaves the strand with the T. The opposite strand is not cleaved. The enzyme also recognizes G/G mismatches and cleaves one strand or the other. TDG enzyme exhibits poor AP endonuclease and AP lyase activities.
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| Applications: |
Mismatch detection
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| Storage: |
Freeze in working aliquots at -20°C to avoid repeated freeze-thawing. The enzyme is supplied in 10 mM HEPES KOH, pH 7.4, 100 mM KCl, 1 mM EDTA, 0.1 mg/ml BSA, and 50% glycerol. |
