MCLAB/T7 Exonuclease/T7G6E-200/10,000 units
Description:T7 Exonuclease is similar to Lambda Exonuclease in that it catalyzes the stepwise hydrolysis of duplex DNA from the 5'-termini, liberating 5'-mononucleotides. However, unlike Lambda Exonuclease, the enzyme has low processivity and it will remove both 5'-hydroxyl and 5'-phosphoryl termini. T7 Exonuclease hydrolyzes duplex DNA non-processively in the 5' -> 3' direction from both 5'-phosphoryl or 5'-hydroxyl nucleotides by liberating oligonucleotides, as well as mononucleotides, until about 50% of the DNA is acid soluble.Source:Purified from an E. coli strain containing a TYB12 intein fusion.Storage Conditions10 mM Tris-HCl5 mM DTT0.1 mM EDTA50% GlycerolpH 8.0 @ 25°CApplication:- Controlled stepwise digestion of double-stranded DNA from the 5'-termini.- Generating ssDNA templates for sequencing via the chain-termination method.Supplied with:10X T7 Exonuclease Buffer10X T7 Exonuclease Buffer:500 mM Potassium Acetate200 mM Tris-acetate100 mM Magnesium Acetate10 mM DTTpH 7.9 @ 25°CUnit Definition:One unit is the amount of enzyme required to release 1 nmol of acid soluble nucleotide in 15 min at 37°C under standard assay conditions.Recommended Storage Condition: -20°CReferences:1. Kerr, C. and Sadowski, P. D. (1972) J. Biol. Chem. 247, 311-318.2. Thomas, K. R. and Olivera, B. M. (1978) J. Biol. Chem. 253, 424-429.3. Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A. and Struhl, K., (1987) Current Protocols in Molecular Biology (John Wiley and Sons, Inc.4. Shon, M., Germino, J. and Bastia, D. (1982) J. Biol. Chem. 257, 13823-13827.5. Nikiforov, T. T., Rendle, R. B., Goelet, P., Rogers, Y. H., Kotewicz,6. M. L., Anderson, S., Trainor, G. L. and Knapp, M. R. (1994) Nucl. Acids Res 22, (20), 4167-4175.7. Kornberg, A. and Baker, T. (1991) DNA Replication, Second Edition, 591. |
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