Technical Data
E9008
Exonuclease III
4000u
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Technical Data |
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E9008 |
Exonuclease III |
4000u |
| Cloning | Storage: -20°CShipping: Blue Ice |
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Source: E.coli cells carrying a cloned E.coli xth gene. Storage Buffer: Supplied as a liquid in 50mM Tris-HCl, pH 8.0, 50mM KCl, 1mM DTT, 50% glycerol. Reaction Buffer (10X) E9008-05: Supplied as a liquid in 660mM Tris-HCl, pH 8.0, 6.6mM MgCl2 Unit Definition: One unit of Exonuclease III catalyzes the release of 1nmol of acid soluble reaction products from E.coli [3H]-DNA in 30 min at 37°C. Applications: Suitable for use in construction of nested unidirectional deletions of DNA fragments, generation of single-stranded template for DNA dideoxy sequencing and site-directed mutagenesis. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. |
Quality Control: Endodeozyribonuclease Assay: No detectable conversion of covalently closed circular DNA to nicked DNA was observed after incubation of 50 units of Exonuclease III with 1ug of pUC19 DNA in 50ul of activity assay for 4 hours at 37ºC. Inhibition and Inactivation: Inhibitiors: Metal chelators, p-chloromercuri benzoate (50-90% inhibitory at 0.1mM) Inactivated by heating at 70°C for 10 min. |
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1. Rogers, S.G., Weiss, B., (1980) Methods Enzymol. 65:201-211. 2. Hoheisel, J.D. (1993) Anal. Biochem. 209:238-246. 3. Henikoff, S., (1984) 28:351-359. 4. Guo, Li-He., Wu, R., (1982) Nucleic Acids Res., 10:2065-2084. 5. Vandeyar, M.A., et al., (1988) Gene 65:129-133. 6. Li, Ch., Evans, R.M., (1997) Nucleic Acids Res. 25:4165-4166. 7. Tomb, J.F., Barcak, G.J., (1989) BioTechniques, 7:932-933.
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