CspA I (Age I isoschizomer)
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CspA I (Age I isoschizomer) Technical BrochureTechnical Brochure
 

CspAI is a restriction enzyme purified from Corynebacterium species.

 

Unit substrate: Lambda DNA.

 

Unit calculation assay conditions: 10 mM Bis Tris Propane-HCl (pH 7.0 @ 25oC), 10 mM MgCl2, 1 mM dithiothreitol, 100 μg/ml bovine serum albumin and DNA. Incubate at 37oC.

 

Absence of contaminants: 50 units of CspA 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 CspA I, greater than 90% of the DNA fragments can be ligated and recut with this enzyme.

 

Storage buffer: 100 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: 65oC for 20 minutes.

 

Methylation Sensitivity:

dam methylation: Not sensitive

dcm methylation: Not sensitive

CpG methylation: Not sensitive

 

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

 

Reagents supplied: 10x UCspAI and 10x K buffer