85-1368-45　Bpu1102I (EspI) 1000U　B2650

特徴

• 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.
• 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 37°C.
• Thermal Inactivation: Enzyme is inactivated by incubation at 80°C for 20min.
• Compatible Ends:Bpu10I, Eco81I, Hpy3I
• Number of Recognition Sites in DNA:Lambda: 6PhiX174: 0M13mp18/19: 0pBR322: 0pUC18/19: 0pUC57: 0pTZ19R/U: 0pBluescriptIIKS(-/+): 0pBluescriptIISK(-/+): 0pACYC177: 0pACYC184: 0