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BoxI (PshAI)

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

FastDigest enzyme available O buffer for 100% activity Optimal incubation at 37°C Cleavage blocked or impaired by CpG methylation Thermal inactivation at 80°C in 20 min Genome qualified LO certified

The BoxI (PshAI) restriction enzyme recognizes GACNN^NNGTC sites and cuts best at 37°C in Tango buffer (Isoschizomers: BstPAI, PshAI)
  
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Lambda DNA digested with BoxI (PshAI)

Lambda DNA digested with BoxI (PshAI), 1% agarose, 7 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

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Conditions for 100% Activity
  • 1X Buffer Tango: 33 mM Tris-acetate (pH 7.9 at 37°C), 10 mM Mg-acetate, 66 mM K-acetate, and 0.1 mg/mL BSA
  • Incubate at 37°C
Digestion of Agarose-embedded DNAMinimum 10 units of the enzyme are required for complete digestion of 1 µg of agarose-embedded lambda DNA in 16 hours.
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
NoteIncubation at 30°C results in a 2.5-fold increase in activity.
Storage BufferBoxI is supplied in: 10 mM potassium phosphate (pH 7.4 at 25°C), 100 mM NaCl, 1 mM EDTA, 1 mM DTT, 0.15% Triton X-100, 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
Tango 37°C 0-20 0-20 0-20 20-50 100 20-50 1X or 2X

Incubation at 30°C results in a 2.5-fold increase in activity.

Methylation Effects

Methylation type Sequence Cleavage effect
CpG 5'...GAm5C G(N)3GTC...3'
3'...CT Gm5C(N)3CAG...5'		 Impaired
CpG 5'...GAm5C GNNm5C GTC...3'
3'...CT Gm5CNN Gm5CAG...5'		 Blocked
CpG 5'...GAC(N)4GTm5C G...3'
3'...CTG(N)4CA Gm5C...5'		 No effect

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
7 1 0 1 0 0
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
0 0 0 0 0 2