| 5'-G T A^T A C-3' |
|
| 3'-C A T^A T G-5' |
|
| Catalog # | B3000-10 |
| Concentration | 10u/ul |
| Source | Bacillus stearothermophilus RFL1107 |
| Buffer | 50mM Tris-HCl, 10mM MgCl2, 100mM NaCl and 0.1mg/ml BSA. Incubate at 37°C |
| Diluent Buffer | 10mM Tris-HCl, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. |
| Storage Buffer | 10mM Tris-HCl, 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol. |
| Storage and Stability | May be stored at 4°C for short-term only. For long-term storage, aliquot and store at -20°C. Aliquots are stable for at least 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. |
| Overdigestion Assay | No detectable change in the specific fragmentation pattern is observed after 320-fold overdigestion (20u/ug lambda DNA x 16 hours) with Bst1107I. |
|
| Ligation/Recutting Assay | After 50-fold overdigestion (3u/ug DNA x 17 hours) with Bst1107I, more than 90% of the pBR322 DNA fragments can be ligated at a 5'-termini concentration of 0.2uM. More than 90% 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 | The mix of pUC57/HindIII, pUC57/PstI and pUC57/Eco32I digests was incubated with 10units of enzyme for 16 hours. After religation and transformation 0.3% of white colonies were detected. |
|
| Methylation Effects | The hemimethylated sequence GTATAm5C is cleaved by Bst1107I at a significantly slower rate. Cleavage is blocked when both strands are methylated at positions: |
| 5'… GTATAm5C…3' |
| 3'…m5CATAT 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 | 3 |
| PhiX174 | 0 |
| M13mp18/19 | 0 |
| pBR322 | 1 |
| pUC18/19 | 0 |
| pUC57 | 0 |
| pTZ19R/U | 0 |
| pBluescriptIIKS(-/+) | 0 |
| pBluescriptIISK(-/+) | 0 |
| pACYC177 | 1 |
| pACYC184 | 1 |
|
| Unit Definition | One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA dam-in 1 hour at 37°C in 50ul of assay buffer. |
