rS. Solfataricus Dpo-4 with Buffer and Cations
rS. Solfataricus Dpo-4 with Buffer and Cations Summary
Dpo4 is a thermophilic Y-family DNA polymerase capable of performing translesion synthesis and allows for coupling to PCR for amplification of damaged DNA. Dpo4 has a molecular weight of 42 kDa and is 95% pure by SDS-PAGE. Translesion synthesis is enhanced by the presence of Mn2+ in the reaction. One unit of enzyme incorporates 1 nmole of dNTP onto the 3’ end of annealed primer in 30 minutes at 60°C.
Assay Conditions and Analysis
1X REC Buffer 15 [10 mM HEPES-NaOH (pH 7.4), 1 mM DTT, 50 mM NaCl, 100 μg/ml BSA, 0.1% Triton X-100], 5 mM MgCl2, 4 pmoles of labeled annealed primer, and serial dilutions of enzyme in a 20 μl reaction volume are incubated for 30 minutes at 60°C. Reaction products are resolved by 20% denaturing polyacrylamide gel electrophoresis, and percent primer extended.
• 10X REC™ Buffer 15
• 100 mM MgCl2
• 50 mM MnCl2
• S. solfataricus Dpo4
For research use only. Not for diagnositic use.
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