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HpaI
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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 M  H  SH  A  K 
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