PauI (BsePI)

Blue Ice

Molecular Biology Grade

5'-G^C G C G C-3'
3'-C G C G C^G-5'

Concentration:
10u/ul

Source:
Paracoccus alcaliphilus ZVK3–3

Buffer:
10mM Tris-HCl (pH 8.5), 10mM MgCl2, 100mM KCl and 0.1mg/ml BSA. Incubate at 37°C

Diluent Buffer:
10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. For longer periods–the Storage Buffer should be used.

Storage Buffer:
10mM Tris-HCl (pH 7.5 at 25°C), 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 50% glycerol.

Overdigestion Assay:
No detectable change in the specific fragmentation pattern is observed after 120-fold overdigestion (7.5u/ug lambda DNA x 16 hours) with PauI.

Ligation Recutting Assay:
After 50-fold overdigestion (3u/ug DNA x 17 hours) with PauI, 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 restriction endonuclease for 4 hours.

Blue White Cloning Assay:
The mix of pUC57/HindIII, pUC57/Eco32I and pUC57/PstI digests was incubated with 10 units of enzyme for 16 hours. After religation and transformation 0.2% of white colonies were detected.

Methylation Effect:
Blocked by CG methylation.

Stability during Prolonged Incubation:
A minimum of 0.2 units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.

Thermal Inactivation:
Enzyme is inactivated by incubation at 80°C for 20min.

Digestion of Agarose-embedded DNA:
A minimum of 5 units of enzyme is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours.

Compatible Ends:
AflIII, AscI, DsaI, MluI

Number of Recognition Sites in DNA:
Lambda: 6
PhiX174: 1
M13mp18/19: 0
pBR322: 0
pUC18/19: 0
pUC57: 0
pTZ19R/U: 0
pBluescriptIIKS(-/+): 2
pBluescriptIISK(-/+): 2
pACYC177: 0
pACYC184: 0

Unit Definition:
One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer.