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  Restriction endonucleases

BseB I (BstN I isoschizomer)

BseB I is a restriction enzyme purified from Bacillus stearothermophilus.

Reagents supplied: 10x R2 buffer

Unit definition: One unit is the amount of enzyme required to digest 1 μg of Lambda DNA in 60 min at 60oC in a total reaction volume of 50 μl.

(SKUs: RE009S, RE009L, REc009S, REc009L)

48.00116.00
  • 10u/μl
  • 40-60u/μl
  • 2000 U
  • 3×2000 U

Product Details

BseB I is a restriction enzyme purified from Bacillus stearothermophilus.

Reagents supplied: 10x R2 buffer

Unit definition: One unit is the amount of enzyme required to digest 1 μg of Lambda DNA in 60 min at 60oC in a total reaction volume of 50 μl.

Digest Guidelines

  • Standard reaction conditions: In order to overcome DNA source variability, a ~10 fold overdigest is suggested:
10U BseB I1μl
10x EQ buffer5μl
DNA substrate1μg
Sterile ultrapure waterUp to 50 μl
  • Speed Digest reaction: Incubate for 15 minutes at 60oC as in standard reaction in 1x R2 buffer (50 mM NaCl, 10 mM Tris-HCl (pH 7.9 @ 25oC), 10 mM MgCl2, 100 μg/ml BSA) or 1x EQ buffer (20 mM Tris-acetate (pH 7.9 @ 25oC), 10 mM Mg-acetate, 50 mM K-acetate, 100 μg/ml B SA).
  • Activity(%) in EnzyQuest buffers -Double digest tips
R1R2R3R4EQ
25-501005025-50

Reactions were performed as for unit definition reaction. The above data could be used as guideline for double digest reactions. Our enzymes are also active and compatible in buffer systems from other vendors.

Heat inactivation: No

Unit calculation assay conditions:

  • 1x R2 buffer
  • Lambda DNA
  • Incubation at 60oC

Storage conditions:

  • 50 mM KCl
  • 10 mM Tris-HCl (pH 7.4)
  • 0.1 mM EDTA
  • 1 mM dithiothreitol
  • 200 μg/ml BSA
  • 50% glycerol
  • Store at -20oC

Non-specific nuclease activity: 500 units of BseB I do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of lambda DNA at 60oC.

Re-ligation and recut: After 10-fold overdigestion with BseB I, less than 50% of the DNA fragments can be ligated and recut with this enzyme.

Note: BseB I-digested DNA is difficult to ligate with T4 DNA Ligase. Ligation is enhanced in the presence of 15% PEG4000.

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