* Size:

HpaI Technical BrochureTechnical Brochure
HpaI is a restriction enzyme purified from a recombinant E.coli strain.


Unit substrate: Lambda DNA.


Unit calculation assay conditions: 50 mM potassium acetate, 20 mM Tris-acetate (pH 7.9 @ 25oC), 10 mM magnesium acetate, 1 mM dithiothreitol, 100 μg/ml bovine serum albumin and DNA. Incubate at 37oC.


Absence of contaminants: 50 units of Hpa I do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of Lambda DNA at 37oC. After 10-fold overdigestion with Hpa I, greater than 95% of the DNA fragments can be ligated and recut with this enzyme.


Storage buffer: 50 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 500 μg/ml bovine serum albumin and 50% glycerol. Store at -20oC.


Heat inactivation: No.


Methylation Sensitivity:

dam methylation: Not sensitive

dcm methylation: Not sensitive

CpG methylation: Blocked by some combinations of overlapping

Star activity: Conditions of high enzyme concentration or glycerol concen-tration >5%, may result in star activity.


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


Reagents supplied: 10x A and 10x K buffer