Nuclease BAL-31 |NEB酶试剂 New England Biolabs

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产品信息

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 
Nuclease BAL-31 |
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.

产品来源

Purified from the culture medium of Alteromonas espejiana BAL-31. Contains a mixture of “fast” and “slow” species of the enzyme (3).

产品类别:
Discontinued (<3 years)

  • 特性和用法

    单位定义

    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.

    反应条件

    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)

    贮存溶液

    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

    热失活

    65°C for 20 minutes

    in the presence of 30mM EGTA

  • 优势和特性

    应用特性

    • Progressive shortening of double-stranded DNA fragments at both termini (4)
    • Restriction site mapping (2)

  • 相关产品

    相关产品

    • t1010-monarch-plasmid-miniprep-kit
    • Monarch® DNA 胶回收试剂盒
    • Monarch® PCR & DNA 纯化试剂盒（5 μg）

  • 注意事项

    1. 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.
    2. If necessary, the enzyme may be diluted in reaction buffer prior to use.
    3. Activity is linear with enzyme concentration.
    4. Heat Inactivation will only work in the presence of 30mM EGTA.

  • 参考文献

    1. Gray, H.B. et al. (1975). Nucl.Acids Res.. 2, 1459-1492.
    2. Legerski, R.J. et al. (1978). Nucl.Acids Res.. 5, 1445-1463.
    3. Wei, C.-F. et al. (1983). J. Biol.Chem.. 258, 13506-13512.
    4. Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual(2nd Ed.). 5.73-5.75.

操作说明、说明书 & 用法

  • 操作说明

    1. Protocol for Nuclease BAL-31

工具 & 资源

  • Web 工具

    • Exo Selector

FAQs & 问题解决指南

  • FAQs

    1. Can Nuclease BAL-31 be used to remove 10 base pairs from the end of a DNA fragment?
    2. Why does all of the DNA get degraded when I use Nuclease BAL-31?
    3. Can Nuclease BAL-31 be heat inactivated?
    4. Is Nuclease BAL-31 active in other NEBuffers?
    5. Can Nuclease BAL-31 treated DNA be cloned?
    6. What is a good control for the BAL-31 nuclease?
    7. Will Nuclease BAL-31 degrade RNA?