|Molecular Biology||Storage: -20°CShipping: Blue Ice|
5'-C T G C A^G-3'
3'-G^A C G T C-5'
Concentration: 10 units/ul (see P9180-05 for 50u/ul)
R1625: Restriction Enzyme Buffer A, 10X
R1625-03: Restriction Enzyme Buffer D, 10X
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.
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.
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.
No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with PstI.
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.
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.
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.
Alw21I, BseSI, Mph1103I, SdaI, SduI
Number of Recognition Sites in DNA:
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.