Technical Data
P9180
PstI 10u/ul
10KU
Molecular Biology Storage: -20CShipping: 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)

Source:
Providencia stuarti

Supplied With:
R1625: Restriction Enzyme Buffer A, 10X
R1625-03: Restriction Enzyme Buffer D, 10X

Concentration:
10u/ul

Incubation Temperature:
37C

Unit Definition:
One unit is defined as the amount of PstI required to digest 1ug of lambda DNA in 1 hour at 37C 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 37C.

Thermal Inactivation:
PstI is not inactivated by incubation at 80C 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 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for 6 months at -20C. 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.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.