| 5'-C C A N N N N N^N T G G-3' |
|
| 3'-G G T N^N N N N N A C C-5' |
|
| Catalog # | B3001 |
| Source | Bacillus stearothermophilus X |
| Concentration | 10u/ul |
| Unit Definition | One unit is defined as the amount of BstXI required to digest 1ug of lambda DNA in 1 hour at 55°C in 50ul of assay buffer |
| Supplied With | R1625: Restriction BstXI Buffer A, 10X: Supplied as a liquid in 33mM Tris-acetate, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA (pH 7.9 at 37°C). |
| R1625-03 | Restriction BstXI Buffer D, 10X: Supplied as a liquid in 50mM Tris-HCl (pH 7.5), 10mM MgCl2, 100mM NaCl and 0.1mg/ml BSA. Incubate at 55°C. Incubation at 37°C results in 50% activity. |
| Diluent and Storage Buffer | Supplied as a liquid in 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. |
| Overdigestion Assay | |
| No detectable change in the specific fragmentation pattern is observed after 15-fold overdigestion (15u/ug lambda DNA x 1 hours) (see Star Activity) |
|
| Ligation/Recutting Assay | |
| After 50-fold overdigestion (3u/ug DNA x 17 hours) with BstXI, 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 10 units of BstXI for 4 hours. |
|
| Blue/White Cloning Assay | |
| The mix of pUC57/HindIII, pUC57/Eco32I and pUC57/PstI digests was incubated with 10 units of BstXI for 16 hours. After religation and transformation 0.1% of white colonies were detected. |
|
| Methylation Effects | |
| Dam | Never overlaps- no effect |
| Dcm | May overlap- blocked |
| CpG | Never overlaps- no effect |
| EcpKl/EcoBl | Never overlaps- no effect |
|
| Star Activity | |
| Excess of BstXI (20u/ug DNA x 1 hour) may result in star activity. |
|
| Stability during Prolonged Incubation | |
| A minimum of 0.1 units of BstXI is required for complete digestion of 1ug of lambda DNA in 16 hours at 55°C. |
|
| Thermal Inactivation | |
| BstXI is inactivated by incubation at 65°C for 20min. |
|
| Digestion of Agarose-embedded DNA | |
| A minimum of 5 units of BstXI is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours. |
|
| Number of Recognition Sites in DNA | |
| Lambda | 13 |
| PhiX174 | 3 |
| pBR322 | 0 |
| pUC57 | 0 |
| pUC18/19 | 0 |
| pTZ19R/U | 0 |
| M13mp18/19 | 0 |
|
| Storage and Stability | |
| May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 12 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 diluent buffer. |
