| Catalog # | B2950 |
| Description | 5'-^G A T C-3' |
| 3'-C T A G^-5' |
| Source | Bacillus species RFL143 |
| Concentration | 10u/ul |
| Unit Definition | One unit is defined as the amount of Bsp143I required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer. |
| Diluent Buffer | For short term storage (3-4 weeks)- dilute with 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. |
| Storage Buffer | For long term storage: 10mM Tris-HCl pH 7.5, 100mM KCl, 1mM DTT, 0.1mM EDTA, 0.15% Triton X-100, 0.2mg/ml BSA and 50% glycerol. |
| Supplied With | R1625-35 Restriction Enzyme Buffer for Bsp143I, 10X: |
| Suplied as a liquid in 33mM Tris-acetate pH 7.9, 10mM magnesium acetate, 0.02% Triton X-100, 0.1mg/ml BSA. |
| R1625 | Restriction Enzyme Buffer A, 10X: |
| Suplied as a liquid in 33mM Tris-acetate pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA. |
| 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 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. |
| |
| Enzyme Properties | |
| Compatible Ends | |
| BamHI, BcII, BgIII, Psul |
|
| Methylation Effects | |
| Dam | Completely overlaps - no effect |
| Dcm | Never overlaps - no effect |
| CpG | May overlaps - blocked |
| EcoKl | Never overlaps - no effect |
| EcoBl | May overlap - no effect |
|
| Stability during Prolonged Incubation | |
| A minimum of 0.1units of Bsp143I is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C. |
|
| Thermal Inactivation | |
| Bsp143I is inactivated by incubation at 65°C for 20min. |
|
| Number of Recognition Sites in DNA | |
| Lambda | 116 |
| M13mp18/19 | 7 |
| pBR322 | 22 |
| pUC18/19, pUC57, pTZ19R/U | 15 |
| X174 | 0 |
| Note | Dpnl, Bsp1431 and Mbol all recognize the same sequence but have different methylation sensitivites and cleavage sites. |
|
| Quality Control | |
| Overdigestion Assay | No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with Bsp143I. |
|
| Ligation/Recutting Assay | After 50-fold overdigestion (3u/ug DNA x 17 hours) with Bsp143I, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 1.2uM. 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 Bsp143I for 4 hours. |
