AluI

One unit is defined as the amount of enzyme required to digest 1µg of λ DNA in 1 hour at 37°C in a total reaction volume of 50µl.

For molecular cloning, restriction site mapping, genotyping, Southern blotting, SNP.

AluI 10,000 units/mlIncludes 10X Cutting Buffer

Liquid. In 100mM KCl, containing 10mM Tris-HCl, pH 7.4, 1mM DTT, 0.1mM EDTA, 50% glycerol and 200µg/ml BSA.

After reconstitution, 1X Cutting Buffer: 50mM potassium acetate, 20mM Tris-acetate, 10mM magnesium acetate, 100µg/ml BSA, pH 7.9, at 25°C.

Exonuclease Activity (Radioactivity Release):
A 50 µl reaction in 1X Cutting Buffer containing 1 µg of a mixture of single and double stranded [³H] E. coli DNA and a minimum of 30 units of AluI incubated for 4 hours at 37°C releases <0.1% of the total radioactivity.

Ligation and Recutting (Terminal Integrity):
After a 10-fold over digestion of λ DNA with AluI, ~75% of the DNA fragments can be ligated with T4 DNA ligase in 16 hours at 16°C. Of these ligated fragments, >95% can be recut with AluI.

Non-Specific DNase Activity (16 Hour):
A 50 µl reaction in 1X Cutting Buffer containing 1 µg of λ DNA and a minimum of 50 Units of AluI incubated for 16 hours at 37°C results in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis.

Reconstitute 10X Cutting Buffer with nuclease-free water.

Produced in an E. coli strain that carries the cloned AluI gene from Arthrobacter luteus.