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

BseJI (BsaBI)

5'...G A T N N↓N N A T C...3'
3'...C T A N N↑N N T A G...5'
Available as a FastDigest enzyme for rapid DNA digestion

0.7% agarose 5 cleavage sites O buffer for 100% activity Star activity Optimal incubation at 65°C Not inactivated at 80°C in 20 min Cleavage blocked or impaired by overlapping Dam methylation LO certified

The BseJI (BsaBI) restriction enzyme recognizes GATNN^NNATC sites and cuts best at 65°C in O buffer (Isoschizomers: BsaBI, Bse8I)
  
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Lambda DNA digested with BseJI (BsaBI)

Lambda DNA digested with BseJI (BsaBI), 1% agarose, 21 cleavage sites

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

  • Molecular cloning
  • Restriction site mapping
  • Genotyping
  • Southern Blot
  • Restriction fragment length polymorphism (RFLP)
  • SNP

Note

  • Incubation at 37°C results in less than 10% activity.
  • Greater than 20-fold overdigestion with BseJI may result in star activity.
  • BseJI is blocked by overlapping dam methylation. To avoid dam methylation, use a dam-, dcm- strain, such as GM2163.

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Conditions for 100% Activity
  • 1X Buffer O: 50 mM Tris-HCl (pH 7.5 at 37°C), 10 mM MgCl2, 100 mM NaCl, and 0.1 mg/mL BSA
  • Incubate at 65°C
  • To ensure higher efficiency of digestion, perform the cleavage reaction under paraffin oil
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage BufferBseJI is supplied in:
10 mM Tris-HCl (pH 7.4 at 25°C), 100 mM NaCl, 1 mM DTT, 1 mM EDTA, 0.2 mg/mL BSA, and 50% (v/v) 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
O (Orange) 65°C NR 100† 100 NR NR 100† 2X†

† Star activity appears at a greater than 5-fold overdigestion (5 units x 1 hour).

Methylation Effects

Methylation type Sequence Cleavage effect
Dam (GATC) 5'...Gm6A TC(N)3ATC...3'
3'...C Tm6AG(N)3TAG...5'		 Blocked
CpG 5'...GAT(N)4ATm5C G...3'
3'...CTA(N)4TA Gm5C...5'		 No effect
CpG 5'...m5C GAT(N)4ATm5C G...3'
3'... Gm5CTA(N)4TA Gm5C...5'		 Not determined

Cleavage efficiency close to the termini of PCR fragments

bp from the recognition site to fragment end
1 2 3 4 5
0 50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
21 2 2 1 0 0
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
0 0 0 0 0 1