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SatI (Fnu4HI)

5'...G C↓N G C...3'
3'...C G N↑C G...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 CpG methylation Thermal inactivation at 65°C in 20 min LO certified

The SatI (Fnu4HI) restriction enzyme recognizes GC^NGC sites and cuts best at 37°C in G buffer (Isoschizomers: Fnu4HI, Fsp4HI, ItaI)
  
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Lambda DNA digested with SatI (Fnu4HI)

Lambda DNA digested with SatI (Fnu4HI), 1.4% agarose, 380 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 Ends
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
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
NoteAt least two copies of SatI (Fnu4HI) recognition site are required for efficient cleavage.
Storage BufferSatI 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
G (Green) 37°C 20-50 100 20-50 20-50 50-100 20-50 1X or 2X

Methylation Effects

  • Dam: never overlaps - no effect.
  • Dcm: never overlaps - no effect.
  • CpG: may overlap - blocked.
  • EcoKI: never overlaps - no effect.
  • EcoBI: never overlaps - no effect.
Methylation type Sequence Cleavage effect
CpG 5'...Gm5C GGC...3'
3'...C Gm5CCG...5'		 Blocked
CpG 5'...GCNGm5C G...3'
3'...CGNC Gm5C...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
380 31 17 42 19 19
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
20 20 23 23 19 37

New sites generated by ligation

Newly generated recognition sites resulting from fill-in of 5'-overhang and self-ligation

Recognition Sequence Newly generated sequence after reaction Enzymes that cleave the newly generated sequence
GC^NGCGCNNGC
  • Cac8I
GC^AGCGCAAGC
  • Cac8I
GC^CGCGCCCGC
  • Cac8I
  • FauI
  • [SsiI (AciI)/FastDigest AciI (SsiI)]
GC^GGCGCGGGC
  • Cac8I
GC^TGCGCTTGC
  • Cac8I