ApaLI
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ApaL I Technical BrochureTechnical Brochure
 

ApaL I is a restriction enzyme purified from Acetobacter pasteurianus (ATCC 12875).

 

Unit substrate: Lambda DNA.

 

Unit calculation assay conditions: 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:  100 units of ApaL 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 ApaL I, 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, 200 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 overlapping

 

Percent Activity in MINOTECH Buffers
L M H SH A K
 100  100  10  <10  10-25 100
 
 

General reaction mixture:

 10U ApaL I  1μl
 10x L or K buffer *  2μl
 DNA substrate  <1μg
 Sterile ultrapure water  Up to 20 μl
Incubate for 15 min at 37oC  
*In the case of L buffer we recommend the addition of 

BSA to a final concentration of 100 μg/ml.

 

 

Reagents supplied: 10x L and 10x K buffer