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BplI

5'...↓ 8(N) G A G (N)5 C T C (N)13↓...3'
3'...↓13(N) C T C (N)5 G A G (N)8 ↓...5'
Available as a FastDigest enzyme for rapid DNA digestion

FastDigest enzyme available Tango buffer for 100% activity Requires SAM for stated activity Optimal incubation at 37°C Thermal inactivation at 80°C in 5 min Genome qualified LO certified

The BplI restriction enzyme recognizes ^(8/13)GAG(N)5CTC(13/8)^ sites and cuts best at 37°C in Tango(+SAM) buffer
  
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Lambda DNA digested with BplI

Lambda DNA digested with BplI, 0.7% agarose, 1 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

BplI requires only Mg2+ for its activity, but is stimulated by S-adenosylmethionine. 0.05 mM S-adenosylmethionine gives more than a 100-fold increase in BplI activity. Still, complete cleavage of some substrates with BplI is difficult to achieve. BplI concentration is determined by the maximum cleavage level achieved when no change in the fragmentation pattern is observed with further enzyme increase.

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Conditions for 100% Activity
  • [1x Buffer Tango] + SAM: [33 mM Tris-acetate (pH 7.9 at 37°C), 10 mM Mg-acetate, 66 mM K-acetate, and 0.1 mg/mL BSA] + 0.05 mM S-adenosylmethionine
  • 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.
Storage BufferBplI is supplied in: 10 mM Tris-HCl (pH 7.4 at 25°C), 100 mM KCl, 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
Tango 37°C 0-20 (+SAM) 20-50 (+SAM) 0-20 (+SAM) 0-20 (+SAM) 100 (+SAM) 20-50 (+SAM) 1X (+SAM)

Methylation Effects

  • Dam: never overlaps - no effect.
  • Dcm: never overlaps - no effect.
  • CpG: may overlap - no effect.
  • EcoKI: may overlap - no effect.
  • EcoKI: never overlaps - no effect.
Methylation type Sequence Cleavage effect
CpG 5'...m5C GAG(N)5CTm5C G...3'
3'... Gm5CTC(N)5GA Gm5C...5'		 No effect

Number of recognition sites in DNA molecules

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