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Nuclease BAL-31

Catalog # Concentration Size List Price Quantity Your Price
M0213S 1000 units/ml 50 units
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Catalog # Size List Price Your Price
M0213S 50 units
Please Inquire
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

  • DNA and RNA nuclease
  • Degrades linear double-stranded DNA from both 5' and 3' termini
  • Acts as an endonuclease that cleaves at nicks, gaps, and single-stranded regions of double-stranded DNA and RNA

Nuclease BAL-31 is ideal for:

  • Progressive shortening of double-stranded DNA
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  • Monarch_MiniprepProtocol_VideoThumb
    What are exonucleases and their applications?
Nuclease BAL-31 exonuclease degrades both 3' and 5' termini of duplex DNA without generating internal scissions. The enzyme is also a highly specific single-stranded endonuclease which cleaves at nicks, gaps and single-stranded regions of duplex DNA and RNA (1,2).
Ligation of Nuclease BAL-31 treated fragments 
Ligation of Nuclease BAL-31 treated fragments
A) Gel electrophoresis of Lambda DNA-HaeIII digest.  
B) Lambda DNA-HaeIII digest after 2 minute incubation with one unit of Nuclease BAL-31. 
C) As in (B) followed by incubation with T4 DNA Ligase.
Product Source
Purified from the culture medium of Alteromonas espejiana BAL-31. Contains a mixture of "fast" and "slow" species of the enzyme (3).
Application Features
  • Progressive shortening of double-stranded DNA fragments at both termini (4)
  • Restriction site mapping (2)

Properties & Usage

Unit Definition

One 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.

Reaction Conditions

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)

Storage Buffer

0.25 mM EDTA
10 mM Tris-HCl
50 mM NaCl
1.5 mM CaCl2
1.5 mM MgCl2
200 µg/ml BSA
50% Glycerol
pH 8 @ 25°C

Heat Inactivation

65°C for 20 minutes

in the presence of 30mM EGTA



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Notes
  • 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 30mM EGTA.
References
  • 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.
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Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety Data Sheets
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