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AdeI (DraIII)

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

FastDigest enzyme available Genome qualified G buffer for 100% activity Optimal incubation at 37°C LO certified Not inactivated at 80°C in 20 min Cleavage blocked or impaired by overlapping CpG methylation Star activity

The AdeI (DraIII) restriction enzyme recognizes CACNNN^GTG sites and cuts best at 37°C in G buffer (Isoschizomers: DraIII)
  
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Lambda DNA digested with AdeI (DraIII)

Lambda DNA digested with AdeI (DraIII), 0.7% agarose, 10 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.

  
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 10 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.
NoteGreater than 20-fold overdigestion with AdeI may result in star activity.
Storage BufferAdeI is supplied in: 10 mM Tris-HCl (pH 7.5 at 25°C), 300 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 0-20 100 20-50 100 100† 20-50 1X† or 2X

† Star activity appears at a greater than 5-fold overdigestion (5 units × 1 hour).

Methylation Effects

  • Dam: never overlaps - no effect.
  • Dcm: never overlaps - no effect.
  • CpG: may overlap - cleavage impaired.
  • EcoKI: may overlap - effect not determined.
  • EcoBI: may overlap - effect not determined.
Methylation type Sequence Cleavage effect
CpG 5'...CAm5C GNNGTG...3'
3'...GT Gm5CNNCAC...5'		 Impaired
CpG 5'...CAm5C GNm5C GTG...3'
3'...GT Gm5CN Gm5CAC...5'		 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
10 1 1 0 0 0
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
1 1 1 1 1 0

New sites generated by ligation

Newly generated recognition sites resulting from removal of 3'-overhang and self-ligation

Recognition Sequence Newly generated sequence after reaction Restriction enzymes that cleave the newly generated recognition sequence
CACNNN^GTGCACGTG
  • Eco72I (PmlI)/FastDigest PmlI (Eco72I)
  • MaeII
  • Ppu21I (BsaAI)/FastDigest BsaAI (Ppu21I)
  • SetI
  • TaiI (MaeII)/FastDigest TaiI