Technical Data
Bpu1102I (EspI)
Molecular Biology Storage: -20CShipping: Blue Ice
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 37C 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 37C.

Dilution/Storage Buffer:: 10mM Tris-HCl, pH 7.4 at 25C, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol.

Quality Control:
Overdigestion Assay:
No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with Bpu1102I.

Ligation/Recutting Assay:
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.

Enzyme Activity:

Methylation Effect:
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 37C.

Thermal Inactivation: Enzyme is inactivated by incubation at 80C for 20min.

Compatible Ends:
Bpu10I, Eco81I, Hpy3I

Number of Recognition Sites in DNA:
Lambda: 6
PhiX174: 0
M13mp18/19: 0
pBR322: 0
pUC18/19: 0
pUC57: 0
pTZ19R/U: 0
pBluescriptIIKS(-/+): 0
pBluescriptIISK(-/+): 0
pACYC177: 0
pACYC184: 0
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.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.