Exonuclease I

For general laboratory use.

Unit Definition: One unit is defined as the amount of enzyme that catalyses the release of 10 nmol acid-soluble nucleotides in a total reaction volume of 50 μl within 30 minutes at 37 °C.

Shipping: shipped on gel packs

Storage Conditions: store at -20 °C
avoid freeze/thaw cycles

Shelf Life: 12 months

Form: liquid

Concentration: 20 units/μl

Applications:

• removal of residual ssDNA, including oligonucleotides, from reaction mixtures
• does not degrade double-stranded DNA or RNA
• requires magnesium and presence of free, accessible 3'-hydroxyl-termini
• active in a wide variety of buffer conditions, allowing for direct addition of enzyme to most reaction mixtures

Description:
Catalyses removal of nucleotides from single-stranded DNA in 3' -> 5' direction.

10x Reaction Buffer:
670 mM Glycine-KOH (pH 9.5 at 25 °C), 100 mM 2-mercaptoethanol and 67 mM MgCl₂.

Assay Conditions:
67 mM Glycin-KOH (pH 9.5 at 25 °C), 10 mM 2-mercaptoethanol, 6.7 mM MgCl₂, 0.17 mg/ml single-stranded [³H]-DNA. Incubation is at 37 °C for 10 min in a reaction volume of 50 μl.

Related products:

• Shrimp Alkaline Phosphatase (rSAP), #EN-174
• SAP-Exo Kit, #PP-218

BIOZ Product Citations:

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Selected References:
Lehman et al. (1964) J. Biol. Cem. 239:2628.
Kushne et al. (1971) Proc. Natl. Acad. Sci. USA 68:824.
Kushner et al. (1972) Proc. Natl. Acad. Sci. USA 69:1366.
Goldmark et al. (1972) J. Biol. Chem. 247:184.
Rosamond et al. (1979) J. Biol. Chem. 254:8646.
Werle et al. (1994) Nuc. Acids Research 22 (20):4354.