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Eco31I (BsaI)

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

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

The Eco31I (BsaI) restriction enzyme recognizes GGTCTC(1/5)^ sites and cuts best at 37°C in G buffer (Isoschizomers: BsaI, Bso31I, BspTNI)
  
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Lambda DNA (dcm-) digested with Eco31I (BsaI)

Lambda DNA (dcm-) digested with Eco31I (BsaI), 0.7% agarose, 2 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

  • Assayed using lambda DNA (dcm-) (#SD0021) as one of the two Eco31I recognition sites in lambda DNA is difficult to cleave. Eco31I cleavage is impaired by overlapping dcm methylation. To avoid dcm 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.
  • Eco31I cleaves downstream of its recognition site and can generate any desired 4 base 5'-overhangs. This feature is useful for direct PCR product cloning.

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Conditions for 100% Activity
  • 1X Buffer G: 10 mM Tris-HCl (pH 7.5 at 37°C), 10 mM MgCl2, 50 mM NaCl, 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 lambda DNA in 16 hours.
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage BufferEco31I is supplied in: 10 mM Tris-HCl (pH 7.5 at 25°C), 200 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
G (Green) 37°C 50-100 100 0-20 0-20 50-100 20-50 1X or 2X

Methylation Effects

Methylation type Sequence Cleavage effect
Dcm (CCWGG) 5'...Cm5CW GGTCTC...3'
3'...G GWm5CCAGAG...5'		 Impaired
CpG 5'...GGTCTm5C G...3'
3'...CCAGA Gm5C...5'		 No effect
CpG 5'...m5C GGTCTC...3'
3'... Gm5CCAGAG...5'		 Impaired

Cleavage efficiency close to the termini of PCR fragments

bp from the recognition site to fragment end
1 2 3 4 5
20-50 50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
2 0 0 1 1 1
pTZ19R/U pTZ57R pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
1 1 1 1 1 0