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

SsiI (AciI)

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

FastDigest enzyme available O buffer for 100% activity Optimal incubation at 37°C Cleavage blocked or impaired by CpG methylation Thermal inactivation at 65°C in 20 min LO certified

The SsiI (AciI) restriction enzyme recognizes CCGC(-3/-1)^ sites and cuts best at 37°C in O buffer (Isoschizomers: AciI, BspACI)
  
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Lambda DNA digested with SsiI (AciI)

Lambda DNA digested with SsiI (AciI), 2% agarose, 516 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 EndsBsp119I, Bsu15I, Hin1I (GR↓CGCC), Hin6I, HpaII, MspI, NarI, Psp1406I, TaqI, XmiI (GT↓CGAC).
Conditions for 100% Activity
  • 1X Buffer O: 50 mM Tris-HCl (pH 7.5 at 37°C), 10 mM MgCl2, 100 mM NaCl, and 0.1 mg/mL BSA
  • Incubate at 37°C
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
NoteLow salt, high glycerol (> 5%) concentrations, pH > 8.0 or a large excess of enzyme may result in star activity.
Storage BufferSsiI 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
O (Orange) 37°C NR 20-50 100 50-100 NR 100 2X

Methylation Effects

Methylation type Sequence Cleavage effect
CpGCCGC Blocked

Cleavage efficiency close to the termini of PCR fragments

bp from the recognition site to fragment end
1 2 3 4 5
50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
516 36 42 67 34 34
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
32 32 36 36 32 56

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
C^CGC
  • BmeT110I* (CY^CGGG)
  • BcnI (NciI)/FastDigest NciI (BcnI)
  • Bme1390I (ScrFI)/FastDigest ScrFI (Bme1390I)
  • BssKI
  • HpaII/FastDigest HpaII
  • MspI (HpaII)/FastDigest MspI
  • Hin1I (BsaHI)/FastDigest BsaHI (Hin1I)* (GR^CGCC)
  • Hin6I (HinP1I)/FastDigest HinP1I (Hin6I) (G^CGC)
  • NarI (GG^CGCC)
  • SsiI (AciI)/FastDigest AciI (SsiI)
  • HpaII/FastDigest HpaII (C^CGG)
  • MspI (HpaII)/FastDigest MspI (C^CGG)
  • HpaII/FastDigest HpaII
  • MspI (HpaII)/FastDigest MspI
G^CGG
  • Hin1I (BsaHI)/FastDigest BsaHI (Hin1I)* (GR^CGCC)
  • Hin6I (HinP1I)/FastDigest HinP1I (Hin6I) (G^CGC)
  • NarI (GG^CGCC)
  • HhaI/FastDigest HhaI
  • Hin6I (HinP1I)/FastDigest HinP1I (Hin6I)

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
C^CGCCCGCGC
  • Bsh1236I (BstUI)/FastDigest Bsh1236I
  • HhaI/FastDigest HhaI
  • Hin6I (HinP1I)/FastDigest HinP1I (Hin6I)
  • [SsiI (AciI)/FastDigest AciI (SsiI)]