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Pst I
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Pst I Technical BrochureTechnical Brochure
PstI is a restriction enzyme purified from a recombinant E.coli strain.

 

Unit substrate: Lambda DNA.

 

Unit calculation assay conditions: 100 mM NaCl, 50 mM Tris-HCl (pH 7.4 @ 25oC), 10 mM MgCl2, 1 mM dithiothreitol, 100 μg/ml bovine serum albumin and DNA. Incubate at 37oC.

 

Absence of contaminants: 200 units of Pst I do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of Lambda DNA at 37oC. After 100-fold overdigestion with Pst I, greater than 95% of the DNA fragments can be ligated and recut with this enzyme.

 

Storage buffer: 200 mM NaCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 200 μg/ml bovine serum albumin, 0.15% Triton X-100 and 50% glycerol. Store at -20oC.

 

Heat inactivation: 80oC for 20 minutes.

 

Methylation Sensitivity:

dam methylation: Not sensitive

dcm methylation: Not sensitive

CpG methylation: Not sensitive



Star activity: Conditions of high enzyme concentration or glycerol concentration>12% may result in star activity.

 

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

 

Reagents supplied: 10x UPstI and 10x K buffer