| G A |
|
| 5'-G T G C C^C-3' |
|
| 3'-C^A C G G G-5' |
|
| C T |
|
| Catalog # | B2918 |
| Concentration | 10u/ul |
| Source | Bacillus stearothermophilus Jo 10–553 |
| Buffer | 10mM Tris-HCl (pH 7.5), 10mM MgCl2, 50mM NaCl and 0.1mg/ml BSA. Incubate at 55°C. (Incubation at 37°C results in less than 20% activity.) |
| Diluent Buffer | 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol, or Storage Buffer. |
| Storage Buffer | 10mM Tris-HCl (pH 7.5 at 25°C), 100mM 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 80-fold overdigestion (5u/ug lambda DNA x 16 hours) with BseSI (see Star Activity). |
|
| Ligation/Recutting Assay | After 50-fold overdigestion (3u/ug DNA x 17 hours) with BseSI, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.1uM. 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. |
|
| Star Activity | A large excess of enzyme (10u/ug DNA x 16 hours) may result in star activity. |
|
| Methylation Effect | Sequence GKGCm5AC is cleaved by BseSI at a significantly slower rate. Overlapping EcoKI 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 55°C. |
|
| Thermal Inactivation | Only small amounts of enzyme (up to 10units) can be inactivated 80°C for 20min. |
|
| Digestion of Agarose-embedded DNA | A minimum of 5units of enzyme is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16 hours. |
|
| Compatible Ends | GGGCC^C–ApaI, Eco24I, SduI GTGCA^C–Alw21I, Mph1103I, PstI, SdaI, SduI |
|
| Number of Recognition Sites in DNA | |
|
| Lambda | 10 |
| PhiX174 | 1 |
| M13mp18/19 | 2* |
| pBR322 | 3 |
| pUC18/19 | 3 |
| pUC57 | 4 |
| pTZ19R/U | 2 |
| pBluescriptIIKS(-/+) | 3 |
| pBluescriptIISK(-/+) | 3 |
| pACYC177 | 2 |
| pACYC184 | 1 |
|
| Unit Definition | One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA in 1 hour at 55°C in 50ul of assay buffer. |
|
| * According to our experimental data, BseSI has only one recognition site in M13mp18/19 DNA at a position 2088. |
