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Eco57I (AcuI)

5'...C T G A A G (N)16...3'
3'...G A C T T C (N)14...5'

Available as a FastDigest enzyme for rapid DNA digestion

FastDigest enzyme available G buffer for 100% activity Requires SAM for stated activity Optimal incubation at 37°C Thermal inactivation at 65°C in 20 min Recombinant enzyme LO certified

The Eco57I (AcuI) restriction enzyme recognizes CTGAAG(16/14)^ sites and cuts best at 37°C in G (+SAM) buffer (Isoschizomers: AcuI)
  
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Lambda DNA digested with Eco57I (AcuI)

Lambda DNA digested with Eco57I (AcuI), 1% agarose, 40 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

  • Eco57I requires only Mg2+ for its activity, but is stimulated by S-adenosylmethionine. 0.01 mM S-adenosylmethionine gives a 100-fold increase in Eco57I activity. Still, complete cleavage of some substrates with Eco57I is difficult to achieve Eco57I concentration is determined by the maximum cleavage level achieved when no change in the fragmentation pattern is observed with further enzyme increase.
  • Low salt, high glycerol (> 5%) concentrations, pH > 8.0 or a large excess of enzyme may result in star activity.
  • Eco57I may remain associated with the cleaved DNA. This may cause DNA band shifting during electrophoresis. To avoid atypical DNA band patterns, use the 6X DNA Loading Dye & SDS Solution for sample preparation or heat the digested DNA in the presence of SDS prior to electrophoresis.

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Conditions for 100% Activity
  • [1X Buffer G] + SAM: [10 mM Tris-HCl (pH 7.5 at 37°C), 10 mM MgCl2, 50 mM NaCl, and 0.1 mg/mL BSA] + 0.01 mM S-adenosylmethionine
  • Incubate at 37°C
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage BufferEco57I is supplied in: 10 mM potassium phosphate (pH 7.4 at 25°C), 100 mM NaCl, 1 mM EDTA, 1 mM DTT, 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) + SAM 37°C 100 (+SAM) 100 (+SAM) 20-50 (+SAM) 20-50 (+SAM) 50-100 (+SAM) 50-100 (+SAM) 1X or 2X (+SAM)

Methylation Effects

  • Dam: never overlaps - no effect.
  • Dcm: never overlaps - no effect.
  • CpG: never overlaps - no effect.
  • EcoKI: never overlaps - no effect.
  • EcoBI: may overlap - effect not determined.

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
40 0 0 2 2 2
pTZ19R/U pTZ57R pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
2 2 2 2 1 2