B1087
BglI
Size
£294.00
- SKU:
- B1087
- Storage Conditions:
- -20°C
- Supplier:
- United States Biological
- Extra Details:
- 5'-G C C N N N N^N G G C-3' 3'-C G G N^N N N N C C G-5' Concentration: 10u/ul Source: Bacillus globigii Buffer: 50mM Tris-HCl (pH 7.5), 10mM MgCl2, 100mM NaCl and 0.1mg/ml BSA Incubate at 37°C. 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 7.5 at 25°C), 300mM NaCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 50% glycerol. Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with BglI. Ligation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with BglI, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.3uM. 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. Methylation Effects: The hemi-methylated sequence GCCN5GGm5C is cleaved by BglI at a significantly slower rate. Cleavage is blocked when both strands are methylated at positions: 5'… GCCN5GGm5C…3' 3'…m5CGGN5CC G…5'. Overlapping CG methylation may influence DNA cleavage. Stability during Prolonged Incubation: A minimum of 0.1units 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 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. Number of Recognition Sites in DNA: Lambda: 29 PhiX174: 0 M13mp18/19: 1 pBR322: 3 pUC18/19: 2 pUC57: 2 pTZ19R/U: 2 pBluescriptIIKS(-/+): 2 pBluescriptIISK(-/+): 2 pACYC177: 1 pACYC184: 2 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.
- Shipping Conditions:
- Blue Ice