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PspP I (Sau96 I isoschizomer)
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PspP I (Sau96 I isoschizomer) Technical BrochureTechnical Brochure
PspPI is a restriction enzyme purified from Psychrobacter immobilis TA137.

 

Unit substrate: Lambda DNA.

 

Unit calculation assay conditions: 50 mM NaCl, 10 mM Tris-HCl (pH 7.9 @ 25oC), 10 mM MgCl2, 1 mM dithiothreitol, 100 μg/ml bovine serum albumin and DNA. Incubate at 25oC.

 

Absence of contaminants: 80 units of PspP I do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of Lambda DNA at 25oC. After 50-fold overdigestion with PspP I, greater than 95% of the DNA fragments can be ligated and recut with this enzyme.

 

Storage buffer: 50 mM NaCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 200 μg/ml bovine serum albumin and 50% glycerol. Store at -20oC.

 

Heat inactivation: 55oC for 15 minutes..

 

Methylation Sensitivity:

dam methylation: Not sensitive

dcm methylation: Blocked by overlapping

CpG methylation: Blocked by overlapping

 

Note: Incubation at 37oC results in 60% activity.

 

Reference: Rina, M., Caufrier, F., Mavromatis, K., Markaki, M., Kokkinidis, M. and Bouriotis, V. (1997) Gene, 197, 353-360.

 

Percent Activity in MINOTECH Buffers
 L M  H  SH  A  K 
50-75  100  50  25-50  10  100 
 
 
General reaction mixture:
10U PspPI 1μl
10x M or K buffer*  2μl 
DNA substrate  <1μg
Sterile ultrapure water  Up to 20 μl
Incubate for 15 min at 37oC  
*In the case of M buffer we recommend the addition of BSA to a final concentration of 100 μg/ml. 

 

Reagents supplied: 10x M and 10x K buffer