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5'-G C^T N A G C-3'
3'-C G A N T^C G-5'
Source: Bacillus pumilus RFL1102
Unit Definition: One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer.
Concentration: 10 units/ul
R1625 Restriction Enzyme Buffer A, 10X,: 33mM Tris-acetate, pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA.
Double Digests: R1625 Restriction Enzyme Buffer A, 10X (Catalog No: R1625) is provided to simplify buffer selection for double digests. Y+/Tangle Buffer is supplied as a liquid in 33mM Tris-acetate, pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA. Incubate at 37°C.
Dilution/Storage Buffer:: 10mM Tris-HCl, pH 7.4 at 25°C, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol.
No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with Bpu1102I.
After 50-fold overdigestion (3u/ug DNA x 17 hours) with Bpu1102I, more than 80% of the DNA fragments can be ligated in 20-40u of T4 DNA ligase/1ug of fragments and 10% PEG at a 5'-termini concentration of 0.2uM. 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 10 units of restriction endonuclease for 4 hours.
Blue/White Cloning Assay:
The mix of pUC57/HindIII, pUC57/Eco32I and pUC57/PstI digests was incubated with 10 units of enzyme for 16 hours. After religation and transformation 0.2% of white colonies were detected.
Bpu1102I cuts GCTNAGm5C.
Stability during Prolonged Incubation:
A minimum of 0.1unit of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Thermal Inactivation: Enzyme is inactivated by incubation at 80°C for 20min.
Bpu10I, Eco81I, Hpy3I
Number of Recognition Sites in DNA:
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.