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BspPI (AlwI)

5'...G G A T C (N)4↓...3'
3'...C C T A G (N)5↑...5'

Tango buffer for 100% activity Optimal incubation at 55°C Cleavage blocked or impaired by Dam methylation Thermal inactivation at 80°C in 20 min LO certified

The BspPI (AlwI) restriction enzyme recognizes GGATC(4/5)^ sites and cuts best at 55°C in Tango buffer (Isoschizomers: AclWI, AlwI)
  
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Lambda DNA digested with BspPI (AlwI)

Lambda DNA digested with BspPI (AlwI), 1.4% agarose, 58 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 30% activity. Assayed using lambda DNA (dam-) (#SD0021). BspPI 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 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 55°C
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage BufferBspPI 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 55°C 20-50 20-50 0-20 0-20 100 0-20 1X

Methylation Effects

Methylation type Sequence Cleavage effect
Dam (GATC)GGm6ATC Blocked
Dcm (CCWGG) 5'...Cm5CW GGATC...3'
3'...G GWm5CCTAG...5'		 No effect
CpG 5'...GGATm5C G...3'
3'...CCTA Gm5C...5'		 No effect
CpG 5'...m5C GGATC...3'
3'... Gm5CCTAG...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
58 0 4 12 10 10
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
10 10 10 10 13 4