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SmoI (SmlI)

5'...CT Y R A G...3'
3'...G A R Y TC...5'

Tango buffer for 100% activity Optimal incubation at 55°C Cleavage blocked or impaired by overlapping Dam methylation Thermal inactivation at 80°C in 20 min Genome qualified LO certified

The SmoI (SmlI) restriction enzyme recognizes C^TYRAG sites and cuts best at 55°C in Tango buffer (Isoschizomers: SmlI)
  
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Lambda DNA digested with SmoI (SmlI)

Lambda DNA digested with SmoI (SmlI), 0.7% agarose, 17 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

Incubation at 37°C results in 10% activity. SmoI cleavage is impaired by overlapping dam methylation. To avoid dam methylation, use a dam-, dcm- strain such as GM2163.Low salt, high glycerol (> 5%) concentrations, pH > 8.0 or a large excess of enzyme may result in star activity.

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Compatible Ends
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 55°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 BufferSmoI is supplied in: 10 mM Tris-HCl (pH 7.5 at 25°C), 50 mM KCl, 1 mM DTT, 0.1 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 55°C 50-100 20-50 0-20 20-50 100 20-50 1X or 2X

Methylation Effects

Methylation type Sequence Cleavage effect
Dam (GATC) 5'...CTYRAGm6A TC...3'
3'...GARYTC Tm6AG...5'
Impaired
CpG 5'...CTm5C GGT...3'
3'...GA Gm5CCA...5'
No effect

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
17 5 5 6 4 4
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
4 4 5 5 4 4

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^TCGAG
  • AbsI (CC^TCGAGG)
  • Eco88I (AvaI)/FastDigest AvaI (Eco88I)* (C^TCGAG)
  • PspXI* (VC^TCGAGC)
  • PspXI* (VC^TCGAGG)
  • PspXI* (VC^TCGAGT)
  • XhoI/FastDigest XhoI (C^TCGAG)
  • BmeT110I
  • Eco88I (AvaI)/FastDigest AvaI (Eco88I)
  • SmoI (SmlI)
  • TaqI/FastDigest TaqI
  • XhoI/FastDigest XhoI
  • SalI/FastDigest SalI (G^TCGAC)
  • TaqI/FastDigest TaqI
  • SgrDI (CG^TCGACG)
  • Hpy99I
  • TaqI/FastDigest TaqI
C^TTAAG
  • BspTI (AflII)/FastDigest AflII (BspTI) (C^TTAAG)
  • BspTI (AflII)/FastDigest AflII (BspTI)
  • FastDigest MseI (SaqAI)
  • SmoI (SmlI)
  • Tru1I (MseI)/FastDigest Tru1I

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^TCGAGCTCGATCGAG
  • Bsh1285I (BsiEI)/FastDigest BsiEI (Bsh1285I)
  • Bsp143I (Sau3AI)/FastDigest Sau3AI (Bsp143I)
  • DpnI/FastDigest DpnI
  • MboI/FastDigest MboI
  • PvuI/FastDigest PvuI
  • [TaqI/FastDigest TaqI]
C^TTAAGCTTAATTAAG
  • PacI/FastDigest PacI (TTAAT^TAA)
  • [FastDigest MseI (SaqAI)]
  • TasI (Tsp509I)/FastDigest Tsp509I (TasI)
  • [Tru1I (MseI)/FastDigest Tru1I]