Progressive shortening of duplex DNA
Inactivation by treatment with EGTA
Supplied with 2X Reaction Buffer
Product SourcePurified from the culture medium of Alteromonas espejiana BAL-31. Contains a mixture of "fast" and "slow" species of the enzyme (3).
The following reagents are supplied with this product:
|Store at (°C)||Concentration|
|Nuclease BAL-31 Reaction Buffer||-20||2X|
Advantages and Features
- Progressive shortening of double-stranded DNA fragments at both termini (4)
- Restriction site mapping (2)
Properties and Usage
Unit DefinitionOne unit is defined as the amount of enzyme required to remove 200 base pairs from each end of linearized double-stranded ΦX174 DNA (40 µg/ml) in a total reaction volume of 50 μl in 10 minutes at 30°C in 1X Nuclease BAL-31 Reaction Buffer.
1X Nuclease BAL-31 Reaction Buffer
Incubate at 30°C
1X Nuclease BAL-31 Reaction Buffer:
20 mM Tris-HCl
600 mM NaCl
12 mM MgCl2
12 mM CaCl2
1 mM EDTA
pH 8 @ 25°C
0.25 mM EDTA
10 mM Tris-HCl
50 mM NaCl
1.5 mM CaCl2
1.5 mM MgCl2
200 μg/ml BSA
pH 8.0 @ 25°C
Heat Inactivation65°C for 10 min
Quality Assurance Statement
- Purified free of detectable double-stranded endonuclease activity.
Material Safety Datasheets
- Duplex products of the exonuclease are a mixture of blunt and staggered ends. This mixture can be cloned directly, although maximal ligation efficiency requires repairing the staggered ends with a suitable DNA polymerase.
- If necessary, the enzyme may be diluted in reaction buffer prior to use.
- Activity is linear with enzyme concentration.
- Heat Inactivation will only work in the presence of 20mM EGTA.
- Gray, H.B. et al. (1975). Nucl. Acids Res.. 2, 1459-1492.
- Legerski, R.J. et al. (1978). Nucl. Acids Res.. 5, 1445-1463.
- Wei, C.-F. et al. (1983). J. Biol. Chem.. 258, 13506-13512.
- Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual (2nd Ed.). 5.73-5.75.
- Can Nuclease BAL-31 be used to remove 10 base pairs from the end of a DNA fragment?
- Why does all of the DNA get degraded when I use Nuclease BAL-31?
- Can Nuclease BAL-31 be heat inactivated?
- Is Nuclease BAL-31 active in other NEBuffers?
- Can Nuclease BAL-31 treated DNA be cloned?
- What is a good control for the BAL-31 nuclease?
- Will Nuclease BAL-31 degrade RNA?