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Home | Molecular Biology | Restriction & Modifying Enzymes | Restriction Enzymes | Conventional

Nb.Bpu10I

5'...C C T N A G C...3'
3'...G G A N T↑C G...5'

R buffer for 100% activity Optimal incubation at 37°C Thermal inactivation at 80°C in 20 min Recombinant enzyme LO certified

The Nb.Bpu10I nicking enzyme recognizes CCTNAGC sites and nicks best at 37°C in R buffer
  
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Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.

Highlights

  • Superior quality – stringent quality control and industry leading manufacturing process
  • Convenient color-coded Five Buffer System
  • Includes universal Tango buffer for double-digestions
  • BSA premixed in reaction buffers
  • Wide selection of restriction endonuclease specificities

Applications

  • Production of single-stranded circular DNA from supercoiled double-stranded plasmids in vitro with subsequent use in DNA sequencing, site-specific mutagenesis, etc
  • Creation of nested deletions
  • Vector preparation for ligation independent cloning method
  • Preparations of covalently closed, double-stranded linear DNA molecules

Note

Nb.Bpu10I may remain associated with the cleaved DNA. This may cause DNA band shifting during electrophoresis. To avoid atypical DNA band patterns, use the 6X DNA Loading Dye & SDS Solution for sample preparation, or heat the digested DNA in the presence of SDS prior to electrophoresis.

  
Storage Condition-20 C
HazardousNo
Conditions for 100% Activity
  • 1X Buffer R: 10 mM Tris-HCl (pH 8.5 at 37°C), 10 mM MgCl2, 100 mM KCl, 0.1 mg/mL BSA
  • Incubate at 37°C
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Nicking and Cleavage
  • Incubation of 10 U of enzyme with 1 µg pUC19 DNA (lacking the recognition sequence of Bpu10I) for 1h at 37°C in 50 µL reaction buffer results in more than 10% conversion to circular form.
  • Incubation of 1 U of enzyme with 1 µg pBR322 DNA for 1h at 37°C in 50 µL reaction buffer results in more than 5% conversion to linear form.
Storage BufferNb.Bpu10I is supplied in: 10 mM Tris-HCl (pH 7.5 at 25°C), 200 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.2 mg/mL BSA, 50% glycerol

Reaction conditions

Recommended buffer for 100% activity Optimal temperature Enzyme activity in Thermo Scientific buffers, % Tango buffer for double digestion
B (blue)
1X		 G (green)
1X		 O (orange)
1X		 R (red)
1X		 Tango
(yellow)
1X / 2X
R (Red) 37°C 0-20 20-50 20-50 100 20-50 50-100 1X or 2X