| 5'-T^G A T C A-3' |
|
| 3'-A C T A G^T-5' |
|
| Catalog # | B0807-01 |
| Concentration | 10u/ul |
| Source | Bacillus caldolyticus |
| 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 50% activity. |
| Double Digests Y+/Tango™ Buffer is provided to simplify buffer selection for double digests. 98% of Fermentas Restriction Endonucleases (REs) work well in a 1X or 2X concentration of the Y+/Tango™ Buffer. Please refer to the Enzyme Activity Chart to choose the best buffer for the two REs in your digest. |
| 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 BclI (see Star Activity). |
|
| Ligation/Recutting Assay | After 50-fold overdigestion (3u/ug DNA x 17 hours) with BclI, more than 90% of the DNA fragments can be ligated at a 5'-termini concentration of 0.1uM. 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. |
|
| Star Activity | A large excess of enzyme (7.5u/ug DNA x 16 hours) may result in star activity. Star activity appears at 10-fold overdigestion (10 units x 1 hour). |
|
| Methylation Effects | BclI cuts neither TGm6ATCA nor TGAThm5CA, but cuts TGATm5CA. |
|
| Blocked by Dam methylation. |
|
| 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 55°C. |
|
| Thermal Inactivation | Only small amounts of enzyme (up to 10units) can be inactivated at 80°C for 20min. |
|
| Compatible Ends | BamHI, BglII, Bsp143I, MboI, PsuI |
|
| Number of Recognition Sites in DNA | |
| Lambda | 8 |
| PhiX174 | 0 |
| M13mp18/19 | 0 |
| pBR322 | 0 |
| pUC18/19 | 0 |
| pUC57 | 0 |
| pTZ19R/U | 0 |
| pBluescriptIIKS(-/+) | 0 |
| pBluescriptIISK(-/+) | 0 |
| pACYC177 | 0 |
| 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 55°C in 50ul of assay buffer. |
