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Hind III
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Hind III Technical BrochureTechnical Brochure
HindIII is a restriction enzyme purified from a recombinant E.coli strain.

 

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 37oC.

 

Absence of contaminants: 700 units of Hind III do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of Lambda DNA at 37oC. After 100-fold overdigestion with Hind III, greater than 98% 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: 65oC for 20 minutes.

 

Methylation Sensitivity:

dam methylation: Not sensitive

dcm methylation: Not sensitive

CpG methylation: Not sensitive

 

Star activity: Star activity may be observed in the presence of Mn2+.

 

Percent Activity in MINOTECH Buffers
 L M  H  SH  A  K 
 25-50 100  10-25  10-25  50  100 
 
 
General reaction mixture:
10U HindIII 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