| 5'-C T G C A^G-3' |
|
| 3'-G^A C G T C-5' |
|
| Catalog # | P9180 |
| Concentration | 10 units/ul (see P9180-05 for 50u/ul) |
| Source | Providencia stuarti |
| Supplied With | R1625: Restriction Enzyme Buffer A, 10X |
| R1625-03: Restriction Enzyme Buffer D, 10X |
| Concentration | 10u/ul |
| Incubation Temperature | 37°C |
| Unit Definition | One unit is defined as the amount of PstI required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer. |
| Diluent Buffer | 10mM Tris-HCl pH 7.4,100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. For longer periods, the Storage Buffer should be used. |
| Storage Buffer | Supplied as a liquid in 10mM Tris-HCl pH 7.4, 200mM sodium chloride, 1mM DTT, 0.1mM EDTA, 0.15% Triton X-100, 0.2mg/ml BSA, 50% glycerol. |
| Enzyme Properties | |
| Overdigestion Assay | |
| No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with PstI. |
|
| Ligation/Recutting Assay | |
| After 50-fold overdigestion (3u/ug DNA x 17 hours) with PstI, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.25uM. 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 PstI for 4 hours. |
|
| Blue/White Cloning Assay | |
| pUC57 was digested at a unique site with 10 units of PstI for 16 hours. After religation and transformation < 1% of white colonies were detected. |
|
| Methylation Effects | |
| Dam/Dcm/CpG/EcoKI/EcoBI | Never overlaps - no effect |
|
| Stability During Prolonged Incubation | |
| A minimum of 0.2 units of PstI is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C. |
|
| Thermal Inactivation | |
| PstI is not inactivated by incubation at 80°C for 20 min. |
|
| Digestion of Agarose-embedded DNA | |
| A minimum of 5 units of PstI is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16 hours. |
|
| Compatible Ends | |
| Alw21I, BseSI, Mph1103I, SdaI, SduI |
|
| Number of Recognition Sites in DNA | |
| Lambda | 28 |
| PhiX174 | 1 |
| pBR322 | 1 |
| pUC57 | 1 |
| pUC18/19 | 1 |
| pTZ19R/U | 1 |
| M13mp18/19 | 1 |
|
| Storage and Stability | |
| May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. 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. |
