| Catalog # | B2940 |
| Sequence | 5'-T T^C G A A-3' |
| 3'-A A G C^T T-5' |
| Concentration | 10u/ul |
| Source | Bacillus species RFL119 |
| Storage and Diluent Buffer | 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. |
| Supplied with | R1625: Supplied as a liquid in 33mM Tris-acetate, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA, pH 7.9 at 37ºC. |
| Methylation Effects | Bsp119I does not cut TTm5CGAA. Blocked by CG methylation. |
| Stability during Prolonged Incubation | A minimum of 0.1units of Bsp119I is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C. |
| Thermal Inactivation | Bsp119I is inactivated by incubation at 80°C for 20min. |
| Number of Recognition Sites in DNA | Lambda: 7 |
| PhiX174, M13mp18/19, pBR322, pUC18/19, pUC57, pTZ19R/U: 0 |
| Digestion of Agarose-embedded DNA | A minimum of 5units of Bsp119I is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours. |
| Unit Definition | One unit is defined as the amount of Bsp119I required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer. |
| Overdigestion Assay | |
| No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with Bsp119I. |
|
| Ligation/Recutting Assay | |
| After 50-fold overdigestion (3u/ug DNA x 17 hours) with Bsp119I, 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 Bsp119I for 4 hours. |
|
| Blue/White Cloning Assay | |
| The mix of pUC57/HindIII, pUC57/PstI and pUC57/Eco32I digests was incubated with 10units of Bsp119I for 16 hours. After religation and transformation 0.5% of white colonies was 1%. |
|
| Compatible Ends | |
| Bsu15I, Hin1I, Hin6I, HpaII, MaeII, MspI, NarI, Psp11406I, TaqI, XmiI |
|
| Protocol for Digestion | |
| Water, nuclease free 16ul |
| R1625 2ul |
| DNA (0.5-1ug/ul) 1ul |
| B2940 0.5-2ul |
| Mix gently and spin down for a few seconds. |
| Incubate at 37°C for 1-16 hours |
|
| Protocol for Digestion of PCR Products Directly After Amplification | |
| PCR Reaction Mixture 10ul (about 1ug of DNA) |
| Water, nuclease free 18ul |
| R1625 2ul |
| B2940 1-2ul |
| Mix gently and spin down for a few seconds. |
| Incubate at 37°C for 1-16 hours |
|
| Storage and Stability | |
| May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. |
