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AarI

5'...C A C C T G C (N)4↓...3'
3'...G T G G A C G (N)8↑...5'

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

The AarI restriction enzyme recognizes CACCTGC(4/8)^ sites and cuts best at 37°C in its own unique (+oligo) buffer
  
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Lambda DNA digested with AarI

Lambda DNA digested with AarI, 1% agarose, 12 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.

For cleavage with AarI at least two copies of its recognition sequence are required. Inclusion of 0.5 µM oligonucleotide with the AarI recognition sequence in the reaction mixture significantly improves cleavage of DNAs, especially of those with a single AarI site.

Still, a complete cleavage of some substrates with AarI is difficult to achieve. Greater than 10-fold overdigestion with AarI may result in star activity. AarI 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.

  
Storage Condition-20 C
HazardousNo
Conditions for 100% Activity
  • [1X Buffer AarI] + oligonucleotide: [10 mM Bis-Tris Propane-HCl (pH 6.5 at 37°C), 10 mM MgCl2, 100 mM KCl, 0.1 mg/mL BSA] + 0.5 µM of oligonucleotide (see Note)
  • Incubate at 37°C
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage BufferAarI is supplied in: 10 mM potassium phosphate (pH7nbsp;7.4 at 25°C), 100 mM KCl, 1 mM EDTA, 1 mM DTT, 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
AarI +oligo Buffer (Unique) 37°C NR (+oligo) NR (+oligo) 0-20 (+oligo) 0-20 (+oligo) NR (+oligo) 50-100 (+oligo) 2X (+oligo)

Methylation Effects

Methylation type Sequence Cleavage effect
CpG 5'...CACCTGm5C G...3'
3'...GTGGAC Gm5C...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
12 0 0 0 0 0
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
0 0 0 0 0 0