| 5'-A^C C G G T-3' |
|
| 3'-T G G C C^A-5' |
|
| Catalog # | B2930 |
| Concentration | 10u/ul |
| Source | Bacillus sphaericus Jo 22–024 |
| 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. |
| Storage Buffer | 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM DTT, 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 BshTI (see Star Activity). |
|
| Ligation/Recutting Assay | After 50-fold overdigestion (3u/ug DNA x 17 hours) with BshTI, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.13uM. 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. |
|
| Blue/White Cloning Assay | The mix of pUC57/HindIII, pUC57/PstI and pUC57/Eco32I digests was incubated with 10units of enzyme for 1 hour. After religation and transformation 0.4% of white colonies were detected. |
|
| Star Activity | A large excess of enzyme (10u/ug DNA x 16 hours) may result in star activity. |
|
| Methylation Effects | BshTI does not cut ACm5CGGT. Blocked by CG methylation. |
|
| 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 20units of enzyme is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours. |
|
| Compatible Ends | Cfr10I, Kpn2I, Cfr9I, Eco88I (C^CCGGG), BsaWI, SgrAI. |
|
| Number of Recognition Sites in DNA | |
|
| Lambda | 13 |
| PhiX174 | 0 |
| M13mp18/19 | 0 |
| pBR322 | 0 |
| pUC18/19 | 0 |
| pUC57 | 0 |
| pTZ19R/U | 0 |
| pBluescriptIIKS(-/+) | 0 |
| pBluescriptIISK(-/+) | 0 |
| pACYC177 | 2 |
| pACYC184 | 4 |
|
| 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. |
