| Catalog # | P1800 |
| Sequence | 5'-G C A T G^C-3' |
| 3'-C^G T A C G-5' |
| Source | Pseudomonas aeruginosa |
| Concentration | 10u/ul |
| Unit Definition | One unit is defined as the amount of PaeI required to digest 1ug of lambda DNA fragments in 1 hr at 37°C in 50ul assay buffer. |
| Diluent Buffer | Supplied as a liquid in 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. |
| Storage Buffer | 10mM potassium phosphate (pH 7.0 at 25°C), 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.5mg/ml BSA, 0.15% Triton X-100 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. |
| R1625-01 | Restriction Enzyme Buffer B, 10X: Supplied as a liquid 10mM Tris-HCl, pH 7.5 at 37°C, 10mM magnesium chloride, 0.1mg/ml BSA. Incubate at 37°C |
| Double Digests | R1625 is provided to simplify buffer selection for double digests. 98% of PaeI, works well in a 1x or 2x. |
| Overdigestion Assay | |
| No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with PaeI. |
|
| Ligation/Recutting Assay | |
| After 50-fold overdigestion (3u/ug DNA x 17 hours) with PaeI, more than 95% 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 PaeI for 4 hours. |
|
| Blue/White Cloning Assay | |
| pUC57 was incubated with 10 units of PaeI for 16 hours. After religation and transformation <1% of white colonies were detected. |
|
| Methylation Effects | |
| Dam | , Dcm, EcoKI: never overlaps-no effect |
| CpG | May overlap- no effect |
| EcoBI | May overlap- blocked |
|
| Stability during Prolonged Incubation | |
| A minimum of 0.1 units of PaeI is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C. |
|
| Thermal Inactivation | |
| PaeI is inactivated by incubation at 65°C for 20min. |
|
| Digestion of Agarose-embedded DNA | |
| A minimum of 5 units of PaeI is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours. |
|
| Number of Recognition Sites in DNA | |
| Lambda 6 |
| pBR322 1 |
| phiX174 0 |
| pUC57, pUC18/19, pTZ19R/U, M13mp18/19 1 |
