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MspC I (Afl II isoschizomer)
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MspC I

(Afl II isoschizomer)

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MspCI is a restriction enzyme purified from Micrococcus species.

 

Unit substrate: Lambda DNA (Hind III digest).

 

Unit calculation assay conditions: 150 mM NaCl, 10 mM Tris-HCl (pH 7.9 @ 25°C), 10 mM MgCl2, 1 mM dithiothreitol, 100 μg/ml BSA. Incubate at 37oC.

 

Absence of contaminants: 80 units of MspCI do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of λ DNA/Hind III digest at 37oC. After 10-fold overdigestion with MspCI, 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 7.9 @ 25°C), 0.1 mM EDTA, 1 mM dithiothreitol, 200 μg/ml BSA 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



Reference: Rina, M., Tzanodaskalaki, M., Karagouni, A., Pagomenou, M. and Bouriotis, V. (1992). Nucleic Acids Res., 20, 1806.

 

Percent Activity in MINOTECH Buffers
 L M  H  SH  A  K 
 <10 25-50  75-100  100  50  100 
 
 
General reaction mixture:
10U MspCI 1μl
10x SH or K buffer*  2μl 
DNA substrate  <1μg
Sterile ultrapure water  Up to 20 μl
Incubate for 15 min at 37oC  
*In the case of SH buffer we recommend the addition of BSA to a final concentration of 100 μg/ml. 

 

Reagents supplied: 10x SH and 10x K buffer