| 5'-C A G^C T G-3' |
|
| 3'-G T C^G A C-5' |
|
| Catalog # | P9390 |
| Source | Proteus vulgaris |
| Unit Definition | One unit is defined as the amount of PvuI required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer. |
| Concentration | 10u/ul |
| Dilution/Storage Buffer | Supplied as a liquid in10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. |
| Supplied With | R1625 Restriction Enzyme Buffer A, 10X |
| Supplied as a liquid in 33mM Tris-acetate, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA, pH 7.9 |
| R1625-02 Restriction Enzyme Buffer C, 10X Supplied as a liquid in 10mM Tris-HCl pH 7.5 10mM Magnesium chloride, 50mM sodium chloride, 0.1mg/ml BSA Inhibition Temperature | 37°C. |
| Storage and Stability | May be stored at 4°C for short-term only. For long-term storage, aliquot and store at -20°C. Stable for at least 12 months |
| Double Digests | |
| Restriction Enzyme Buffer A is provided to simplify buffer selection for double digests. 98% of Restriction Endonucleased work well in a 1x or 2x concentration of theRestriction Enzyme Buffer A. |
|
| Number of Recognition Sites | |
| Lambda | 15 |
| M13mp18/19 | 3 pBR322: 1 |
| pUC18/19 | 2 pUC57 : 2 |
| pTZ19R/U | 2 |
| X174 | 0 |
|
| Thermal Inactivation | |
| PvuI is inactivated by incubation at 80°C for 20min. |
|
| Overdigestion Assay | No detectable change in the specific fragmentation pattern is observed after 10-fold overdigestion (15u/ug lambda DNA x 1 hours) with PvuII (see Star Activity). |
|
| Ligation/Recutting Assay | After 10-fold overdigestion (5u/ug DNA x 2 hours) with PvuII, more than 90% of the DNA fragments can be ligated at a 5'-termini concentration of 0.06uM. 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 PvuI for 4 hours. |
|
| Star Activity | A large excess of enzyme (15u/ug DNA x 1 hours) may result in star activity. Star activity appears at 10-fold overdigestion (10 units x 1 hour). |
|
| Methylation Effect | Dam |
| EcoBI | May overlaps- not determined |
|
| Stability during Prolonged Incubation | A minimum of 0.2units of PvuI are required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C. |
