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Esp3I (BsmBI)

5'...C G T C T C (N)1↓...3'
3'...G C A G A G (N)5↑...5'
Available as a FastDigest enzyme for rapid DNA digestion

FastDigest enzyme available Tango buffer for 100% activity Requires DTT for stated activity Optimal incubation at 37°C Cleavage blocked or impaired by CpG methylation Thermal inactivation at 65°C in 20 min Genome qualified Recombinant enzyme LO certified

The Esp3I (BsmBI) restriction enzyme recognizes CGTCTC(1/5)^ sites and cuts best at 37°C in Tango (+DTT) buffer (Isoschizomers: BsmBI)
  
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Lambda DNA digested with Esp3I (BsmBI)

Lambda DNA digested with Esp3I (BsmBI), 1% agarose, 14 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

The enzyme requires DTT. Freshly made DTT should be added to the reaction buffer. Esp3I cleaves downstream of its recognition site and can generate any desired 4 base 5'-overhangs. This feature is useful for PCR product cloning.

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Conditions for 100% Activity
  • [1X Buffer Tango] + DTT: [33 mM Tris-acetate (pH 7.9 at 37°C), 10 mM Mg-acetate, 66 mM K-acetate, and 0.1 mg/mL BSA] + 1.0 mM DTT
  • 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 lambda DNA in 16 hours.
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage BufferEsp3I is supplied in: 10 mM potassium phosphate (pH 7.4 at 25°C), 100 mM KCl, 1 mM EDTA, 7 mM 2-mercaptoethanol, 0.5 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 100 (+DTT) 20-50 (+DTT) 0-20 (+DTT) 0-20 (+DTT) 100 (+DTT) 0-20 (+DTT) 1X (+DTT)

Methylation Effects

  • Dam: never overlaps - no effect.
  • Dcm: never overlaps - no effect.
  • CpG: completely overlaps - blocked.
  • EcoKI: never overlaps - no effect.
  • EcoBI: may overlap - effect not determined.
Methylation type Sequence Cleavage effect
CpGCGTCTC 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
14 0 1 1 2 2
pTZ19R/U pTZ57R pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
0 0 0 0 1 2