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SmaI

5'...C C CG G G...3'
3'...G G GC C C...5'

Available as a FastDigest enzyme for rapid DNA digestion

FastDigest enzyme available Tango buffer for 100% activity Optimal incubation at 30°C Cleavage blocked or impaired by CpG methylation Thermal inactivation at 65°C in 20 min High concentration available Genome qualified Recombinant enzyme LO certified

The SmaI restriction enzyme recognizes CCC^GGG sites and cuts best at 30°C in Tango buffer (Isoschizomers: TspMI, XmaCI; neoschizomer: XmaI)
  
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Lambda DNA digested with SmaI

Lambda DNA digested with SmaI, 0.7% agarose, 3 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.

  
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 30°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.
Note
  • Incubation at 25°C results in 100% activity.
  • Incubation at 37°C results in 50% activity.
  • SmaI has a half-life of 15 min at 37°C.
  • SmaI needs K+ to work for activity.
Storage BufferSmaI 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.
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
Tango 30°C 50-100 0-20 0-20 0-20 100 0-20 1X

Methylation Effects

Methylation type Sequence Cleavage effect
CpGCCCGGG 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
3 0 1 0 1 1
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
1 1 1 1 1 0

New sites generated by ligation

Newly generated recognition sites resulting from ligation of blunt DNA ends

Recognition sequence Second restriction enzyme Restriction enzymes cleaving the newly generated recognition sequence
CCC^GGG
  • DpnI/FastDigest DpnI (GA^TC)
  • Ecl136II (EcoICRI)/FastDigest Ecl136II (GAG^CTC)
  • MbiI (BsrBI)/FastDigest BsrBI (MbiI)* (CCG^CTC)
  • MnlI/FastDigest MnlI
  • Eco32I (EcoRV)/FastDigest EcoRV (Eco32I) (GAT^ATC)
  • BccI
  • Eco47III (AfeI)/FastDigest AfeI (Eco47III) (AGC^GCT)
  • EheI (SfoI)/FastDigest EheI (GGC^GCC)
  • FspAI/FastDigest FspAI* (RTGC^GCAC)
  • FspAI/FastDigest FspAI* (RTGC^GCAT)
  • NsbI (FspI)/FastDigest FspI (NsbI) (TGC^GCA)
  • FauI
  • SsiI (AciI)/FastDigest AciI (SsiI)
  • MbiI (BsrBI)/FastDigest BsrBI (MbiI)* (GAG^CGG)
  • MspA1I* (CMG^CGG)
  • BcnI (NciI)/FastDigest NciI (BcnI)
  • Bme1390I (ScrFI)/FastDigest ScrFI (Bme1390I)
  • BseDI (BsaJI)/FastDigest BsaJI (BseDI)
  • BssKI
  • HpaII/FastDigest HpaII
  • MspI (HpaII)/FastDigest MspI
  • PdiI (NaeI)/FastDigest NaeI (PdiI) (GCC^GGC)
  • BcnI (NciI)/FastDigest NciI (BcnI)
  • Bme1390I (ScrFI)/FastDigest ScrFI (Bme1390I)
  • BssKI
  • HpaII/FastDigest HpaII
  • MspI (HpaII)/FastDigest MspI
  • SrfI (GCCC^GGGC)
  • BcnI (NciI)/FastDigest NciI (BcnI)
  • Bme1390I (ScrFI)/FastDigest ScrFI (Bme1390I)
  • BmeT110I
  • BseDI (BsaJI)/FastDigest BsaJI (BseDI)
  • BssKI
  • Cfr9I (XmaI)
  • Eco88I (AvaI)/FastDigest AvaI (Eco88I)
  • HpaII/FastDigest HpaII
  • MspI (HpaII)/FastDigest MspI
  • SmaI/FastDigest SmaI

References

Citations