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AluI

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

FastDigest enzyme available Optimal incubation at 37°C LO certified Tango buffer for 100% activity Thermal inactivation at 65°C in 20 min

The AluI restriction enzyme recognizes AG^CT sites and cuts best at 37°C in Tango buffer (Isoschizomers: AluBI)
  
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Lambda DNA digested with AluI, 1.4% agarose, 143 cleavage sites

Lambda DNA digested with AluI
1.4% agarose
143 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 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 37°C
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage BufferAluI is supplied in 10 mM Tris-HCl (pH 7.5 at 25°C), 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.15% Triton X-100, 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
Tango 37°C 50-100 0-20 0-20 0-20 100 20-50 1X or 2X

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 - blocked.
Methylation type Sequence Cleavage effect
EcoBI (TGA(N)8TGCT) 5'...TGm6AGCT(N)5 TGCT...3'
3'...AC TCGA(N)5m6ACGA...5'		 Blocked

Cleavage efficiency close to the termini of PCR fragments

bp from the recognition site to fragment end
1 2 3 4 5
0-20 20-50 50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
143 24 27 17 16 16
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
17 17 17 17 12 13

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
AG^CT
  • AjiI (BmgBI)* (CAC^GTC)
  • AjiI (BmgBI)* (GAC^GTG)
  • Eco105I (SnaBI)/FastDigest SnaBI (Eco105I) (TAC^GTA)
  • Eco72I (PmlI)/FastDigest PmlI (Eco72I) (CAC^GTG)
  • Ppu21I (BsaAI)/FastDigest BsaAI (Ppu21I)* (YAC^GTA)
  • Ppu21I (BsaAI)/FastDigest BsaAI (Ppu21I)* (YAC^GTG)
  • ZraI (GAC^GTC)
  • SetI
  • Bst1107I (BstZ17I)/FastDigest BstZ17I (Bst1107I) (GTA^TAC)
  • Csp6I (CviQI)/FastDigest Csp6I
  • RsaI/FastDigest RsaI
  • BsuRI (HaeIII)/FastDigest HaeIII (BsuRI) (GG^CC)
  • CviJI* (RG^CC)
  • Eco147I (StuI)/FastDigest StuI (Eco147I) (AGG^CCT)
  • Eco47III (AfeI)/FastDigest AfeI (Eco47III) (AGC^GCT)
  • MlsI (MscI)/FastDigest MscI (MlsI) (TGG^CCA)
  • CviJI
  • CviJI* (RG^CT)
  • Ecl136II (EcoICRI)/FastDigest Ecl136II (GAG^CTC)
  • MbiI (BsrBI)/FastDigest BsrBI (MbiI)* (CCG^CTC)
  • MspA1I* (CMG^CTG)
  • PvuII/FastDigest PvuII (CAG^CTG)
  • AluI/FastDigest AluI
  • CviJI
  • SetI
  • Eco32I (EcoRV)/FastDigest EcoRV (Eco32I) (GAT^ATC)
  • Bsp143I (Sau3AI)/FastDigest Sau3AI (Bsp143I)
  • DpnI/FastDigest DpnI
  • MboI/FastDigest MboI
  • EheI (SfoI)/FastDigest EheI (GGC^GCC)
  • BsuRI (HaeIII)/FastDigest HaeIII (BsuRI)
  • CviJI