Exonuclease I, E. coli (ExoI)

The Exonuclease I (ExoI) degrades single-stranded DNA in a 3'=>5' direction, releasing deoxyribonucleoside 5'-monophosphates in a stepwise manner and leaving 5'-terminal dinucleotides intact. It does not cleave DNA strands with terminal 3'-OH groups blocked by phosphoryl or acetyl groups (1).\n\nSource: \nE. coli cells with a cloned E. coli sbcB gene.\n\nConcentration: ~20u/ul\n\nUnit Definition:\nOne unit of E9007-85C catalyzes the release of 10 nano-moles of acid soluble nucleotides in 30 min at 37C.\n\nActivity Assay:\n67mM glycine-KOH pH 9.5, 6.7mM MgCl2, 1mM DTT, 0.17mg/ml single stranded [3H]-DNA.\n\nForm:\nSupplied as a liquid in 20mM Tris-HCl, pH 7.5, 0.1mM EDTA, 1mM DTT and 50% glycerol.\n\nSupplied with:\nE9007-85C1: 10X Reaction Buffer: Supplied as a liquid in 670mM glycine-KOH, pH 9.5 at 25 degrees C, 67mM MgCl2, 10mM DTT.\n\nApplications:\nSuitable for use in the purification of PCR products, removal of single-stranded DNA containing a 3'-hydroxyl terminus from nucleic acid mixtures and assay for the presence of single-stranded DNA with a 3'-hydroxyl terminus. Other applications not tested.\n\nProtocol for Elimination of Oligonucleotides and dNTPs from PCT mixtures:\n1. Remove a 5ul aliquot from a reaction mixture.\n2. Add 10u of E9007-85C and 2u of Shrimp Alkaline Phosphatase, P4071-05.\n3. Mix and incubate at 37C for 15 mion.\n4. Inactivate the enzymes by heating at 85C for 15 min.\n*Note: After inactivation, the aliquot can be used directly for DNa sequencing. The Exonuclease I is not suitable for removing 3

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SPECIFICATIONS

Catalog Number

E9007-85C

Size

4000U

References

Lehman, I.R., Nussbaum, A.L., J. Biol. Chem., 239, 2628-2636, 1964. Werle, E., et al., Nucleic Acids Res., 22, 4354-4355, 1994. Nabavi S., Nazar R.N., Anal Biochem., 2005, 2, 346-348, 2005. Rosamond, J., et al., J. Biol. Chem., 254, 8646-8652, 1979.

Alternative Names

Supplied with 10X Reaction Buffer