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Psp5II (PpuMI)

5'...R G↓G W C C Y...3'
3'...Y C C W G↑G R...5'
Available as a FastDigest enzyme for rapid DNA digestion

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

The Psp5II (PpuMI) restriction enzyme recognizes RG^GWCCY sites and cuts best at 37°C in G buffer (Isoschizomers: PpuMI, PspPPI)
  
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Lambda DNA digested with Psp5II (PpuMI)

Lambda DNA digested with Psp5II (PpuMI), 0.7% agarose, 3 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
Compatible EndsCfr13I, CpoI, Eco47I, SanDI.
Conditions for 100% Activity
  • 1X Buffer G: 10 mM Tris-HCl (pH 7.5 at 37°C), 10 mM MgCl2, 50 mM NaCl, and 0.1 mg/mL BSA
  • Incubate at 37°C
Digestion of Agarose-embedded DNAMinimum 5 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 BufferPsp5II is supplied in: 10 mM Tris-HCl (pH 7.4 at 25°C), 100 mM KCl, 1 mM DTT, 1 mM EDTA, 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
G (Green) 37°C 0-20 100 20-50 20-50 50-100 100 1X or 2X

Methylation Effects

  • Dam: never overlaps - no effect.
  • Dcm: may overlap - blocked.
  • CpG: never overlaps - no effect.
  • EcoKI: never overlaps - no effect.
  • EcoBI: may overlap - effect not determined.
Methylation type Sequence Cleavage effect
Dcm (CCWGG) 5'...RGGWCm5CT GG...3' 3'...YCCWG GAm5CC...5' Blocked

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
3 0 0 2 0 0
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
0 0 0 0 0 2

New sites generated by ligation

Newly generated recognition sites resulting from ligation of protruding compatible DNA ends

Recognition Sequence Second Enzyme Enzymes recognizing newly generated recognition sequence
AG^GACCY
  • CpoI (RsrII)/FastDigest RsrII (CpoI)* (CG^GACCG)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)* (G^GACC)
  • BmgT120I
  • Cfr13I (Sau96I)/FastDigest Sau96I (Cfr13I)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)
  • FastDigest SanDI (KflI)* (GG^GACCC)
  • BmgT120I
  • BspLI (NlaIV)/FastDigest NlaIV (BspLI)
  • Cfr13I (Sau96I)/FastDigest Sau96I (Cfr13I)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)
  • EcoO109I (DraII)/FastDigest EcoO109I
  • Psp5II (PpuMI)/FastDigest PpuMI (Psp5II)
AG^GTCCY
  • CpoI (RsrII)/FastDigest RsrII (CpoI)* (CG^GTCCG)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)* (G^GTCC)
  • BmgT120I
  • Cfr13I (Sau96I)/FastDigest Sau96I (Cfr13I)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)
  • SetI
  • FastDigest SanDI (KflI)* (GG^GTCCC)
  • BmgT120I
  • BspLI (NlaIV)/FastDigest NlaIV (BspLI)
  • Cfr13I (Sau96I)/FastDigest Sau96I (Cfr13I)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)
  • EcoO109I (DraII)/FastDigest EcoO109I
  • Psp5II (PpuMI)/FastDigest PpuMI (Psp5II)
  • SetI
GG^GACCY
  • CpoI (RsrII)/FastDigest RsrII (CpoI)* (CG^GACCG)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)* (G^GACC)
  • BmgT120I
  • BspLI (NlaIV)/FastDigest NlaIV (BspLI)
  • Cfr13I (Sau96I)/FastDigest Sau96I (Cfr13I)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)
  • FaqI (BsmFI)/FastDigest BsmFI (FaqI)
  • FastDigest SanDI (KflI)* (GG^GACCC)
  • BmgT120I
  • BspLI (NlaIV)/FastDigest NlaIV (BspLI)
  • Cfr13I (Sau96I)/FastDigest Sau96I (Cfr13I)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)
  • EcoO109I (DraII)/FastDigest EcoO109I
  • FaqI (BsmFI)/FastDigest BsmFI (FaqI)
  • FastDigest SanDI (KflI)
  • Psp5II (PpuMI)/FastDigest PpuMI (Psp5II)
GG^GTCCY
  • CpoI (RsrII)/FastDigest RsrII (CpoI)* (CG^GTCCG)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)* (G^GTCC)
  • BmgT120I
  • BspLI (NlaIV)/FastDigest NlaIV (BspLI)
  • Cfr13I (Sau96I)/FastDigest Sau96I (Cfr13I)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)
  • FastDigest SanDI (KflI)* (GG^GTCCC)
  • BmgT120I
  • BspLI (NlaIV)/FastDigest NlaIV (BspLI)
  • Cfr13I (Sau96I)/FastDigest Sau96I (Cfr13I)
  • Eco47I (AvaII)/FastDigest AvaII (Eco47I)
  • EcoO109I (DraII)/FastDigest EcoO109I
  • FastDigest SanDI (KflI)
  • Psp5II (PpuMI)/FastDigest PpuMI (Psp5II)