| 5'-A G G^C C T-3' |
|
| 3'-T C C^G G A-5' |
|
| Catalog # | E0374-65 |
| Concentration | 10u/ul |
| Source: E.coli that carries the cloned eco147IR gene from E.coli RFL147 |
| Restriction Enzyme Buffer A, 10X | (R1625)33mM Tris-acetate, pH 7.9, 10mM magnesium acetate, 66mM potassium acetate and 0.1mg/ml BSA. Incubate at 37°C. Supplied to simplify buffer selection for double digests. |
| Restriction Enzyme Buffer B, 10X | (R1625-01): |
| 10mM Tris-HCl (pH 7.5), 10mM MgCl2 and 0.1mg/ml BSA. Incubate at 37°C |
| 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 320-fold overdigestion (20u/ug lambda DNA x 16 hours) with Eco147I. |
|
| Ligation/Recutting Assay | |
| After 50-fold overdigestion (3u/ug DNA x 17 hours) with Eco147I, more than 90% of the DNA fragments can be ligated at a 5'-termini concentration of 0.03uM. 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 10 units of restriction endonuclease for 4 hours. |
|
| Methylation Effects | |
| Blocked by overlapping Dcm methylation. |
|
| Blue/White Cloning Assay | |
| pUC57 was digested at a unique site with 10 units of enzyme for 16 hours. After religation and transformation 0.7% of white colonies were detected |
|
| Stability during Prolonged Incubation | A minimum of 0.1 units 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 80°C for 20min. |
|
| Digestion of Agarose-embedded DNA | A minimum of 5 units of enzyme is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours. |
|
| Number of Recognition Sites in DNA | |
| Lambda | 6 |
| PhiX174, pUC57 | 1 |
| M13mp18/19, pBR322, pUC18/19, pTZ19R/U | 0 |
