Recently Viewed

Home | Molecular Biology | Restriction & Modifying Enzymes | Restriction Enzymes | Conventional

SfaAI (AsiSI)

5'...G C G A T↓C G C...3'
3'...C G C↑T A G C G...5'
Available as a FastDigest enzyme for rapid DNA digestion

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

The SfaAI (AsiSI) restriction enzyme recognizes GCGAT^CGC sites and cuts best at 37°C in Tango buffer (Isoschizomers: AsiSI, RgaI, SgfI)
  
Loading...
Ad2 DNA digested with SfaAI (AsiSI)

Ad2 DNA digested with SfaAI (AsiSI), 1% agarose, 1 cleavage site

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 EndsBsh1285I, PacI, PvuI.
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 37°C
Digestion of Agarose-embedded DNAMinimum 5 units of the enzyme are required for complete digestion of 1 µg of agarose-embedded pJET1 DNA with inserted SfaAI recognition site in 16 hours.
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
NoteAssayed using linearized pJET1 DNA with inserted SfaAI recognition site.
Storage BufferSfaAI 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 50-100 0-20 0-20 0-20 100 0-20 1X

Methylation Effects

Methylation type Sequence Cleavage effect
Dam (GATC)GCGm6ATCGC No effect
CpGGCGATCGC 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

Number of recognition sites in DNA molecules

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

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
GCGAT^CGC
  • Bsh1285I (BsiEI)/FastDigest BsiEI (Bsh1285I)* (CGAT^CG)
  • PvuI/FastDigest PvuI (CGAT^CG)
  • Bsh1285I (BsiEI)/FastDigest BsiEI (Bsh1285I)
  • Bsp143I (Sau3AI)/FastDigest Sau3AI (Bsp143I)
  • DpnI/FastDigest DpnI
  • MboI/FastDigest MboI
  • PvuI/FastDigest PvuI
  • PacI/FastDigest PacI (TTAAT^TAA)
  • FastDigest MseI (SaqAI)
  • Tru1I (MseI)/FastDigest Tru1I

Newly generated recognition sites resulting from removal of 3'-overhang and self-ligation

Recognition Sequence Newly generated sequence after reaction Restriction enzymes that cleave the newly generated recognition sequence
GCGAT^CGCGCGCGC
  • Bsh1236I (BstUI)/FastDigest Bsh1236I
  • Cac8I
  • HhaI/FastDigest HhaI
  • Hin6I (HinP1I)/FastDigest HinP1I (Hin6I)
  • PauI (BssHII)/FastDigest BssHII (PteI)