|Molecular Biology||Storage: -20°CShipping: Blue Ice|
5'-G G G C C^C-3'
3'-C^C C G G G-5'
Source: Acetobacter pasteurianus
Buffer: 10mM Tris-HCl (pH 7.5), 10mM MgCl2 and 0.1mg/ml BSA. Incubate at 37°C. (Incubation at 30°C results in a 2-fold increase in activity.)
Diluent Buffer: 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. For longer periods–the Storage Buffer should be used.
Storage Buffer: 10mM Tris-HCl (pH 7.4 at 25°C), 50mM NaCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol.
Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 320-fold overdigestion (20u/ug lambda DNA x 16 hours) with ApaI.
Ligation/Recutting Assay After 50-fold overdigestion (3u/ug DNA x 17 hours) with ApaI, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.02uM. More than 95% of these can be recut.
Labeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10units of restriction endonuclease for 4 hours.
Blue/White Cloning Assay pUC57 was digested at a unique site with 10units of enzyme for 16 hours. After religation and transformation 0.3% of white colonies were detected.
Methylation Effects: Sequence GGGCCm5C is cleaved approximately 150 times slower than the unmethylated one. Overlapping Dcm or CG methylation may influence DNA cleavage.
Stability during Prolonged Incubation: A minimum of 0.2units of enzyme is required for complete digestion of 1ug of DNA in 16 hours at 37°C.
Thermal Inactivation: Enzyme is inactivated by incubation at 65°C for 20min.
Digestion of Agarose-embedded DNA: A minimum of 5units of enzyme is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours.
Compatible Ends: GGGCC^C–BseSI, GGGCC^C–Eco24I, GGGCC^C–SduI
Number of Recognition Sites in DNA:
Unit Definition: One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA-CpoI fragments in 1 hour at 37°C in 50ul of assay buffer.
Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.