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PaeI (SphI)

5'...G C A T GC...3'
3'...CG T A C G...5'

Available as a FastDigest enzyme for rapid DNA digestion

FastDigest enzyme available B buffer for 100% activity Optimal incubation at 37°C Thermal inactivation at 65°C in 20 min Genome qualified LO certified

The PaeI (SphI) restriction enzyme recognizes GCATG^C sites and cuts best at 37°C in B buffer (Isoschizomers: BbuI, SphI)
  
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Lambda DNA digested with PaeI (SphI)

Lambda DNA digested with PaeI (SphI), 0.7% agarose, 6 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.

  
Compatible EndsHin1II, XceI.
Conditions for 100% Activity
  • 1X Buffer B: 10 mM Tris-HCl (pH 7.5 at 37°C), 10 mM MgCl2, 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.
HazardousNo
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage BufferPaeI is supplied in: 10 mM potassium-phosphate (pH 7.0 at 25°C), 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.15% Triton X-100, 0.5 mg/mL BSA, and 50% (v/v) glycerol.
Storage Condition-20 C

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
B (Blue) 37°C 100 50-100 0-20 0-20 50-100 0-20 1X

Methylation Effects

Methylation type Sequence Cleavage effect
CpG 5'...m5C GCATGm5C G...3'
3'... Gm5CGTAC Gm5C...5'
No effect
EcoBI (TGA(N)8TGCT) 5'...TGm6A(N)5GCA TGCT...3'
3'...AC T(N)5CGTm6ACGA...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 0-20 20-50 50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
6 0 1 1 1 1
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
1 1 0 0 0 1

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
GCATG^C
  • Hin1II (NlaIII)/FastDigest NlaIII (Hin1II) (CATG)
  • CviAII
  • FaiI
  • FatI
  • Hin1II (NlaIII)/FastDigest NlaIII (Hin1II)
  • XceI (NspI)/FastDigest NspI (XceI)* (RCATG^C)
  • Cac8I
  • CviAII
  • FaiI
  • FatI
  • Hin1II (NlaIII)/FastDigest NlaIII (Hin1II)
  • PaeI (SphI)/FastDigest SphI (PaeI)
  • XceI (NspI)/FastDigest NspI (XceI)
  • XceI (NspI)/FastDigest NspI (XceI)* (RCATG^T)
  • CviAII
  • FaiI
  • FatI
  • Hin1II (NlaIII)/FastDigest NlaIII (Hin1II)
  • XceI (NspI)/FastDigest NspI (XceI)

References

Citations