Source: Micrococcus species R9
Assayed on: Lambda DNA (dcm-)
Unit definition: One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (dcm-) in 1 hour at 37°C in a total reaction volume of 50 μl.
Optimal SE-buffer: O (50 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
Activity in SEBuffers:
SEBuffer B 50 - 50%
SEBuffer G 50 - 75%
SEBuffer O 100%
SEBuffer W 50 - 75 %
SEBuffer Y 50 - 75%
SEBuffer ROSE 100%
When using a buffer other than the optimal (suppied) SEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
Optimal temperature: 37°C
Storage conditions: 10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0,1 mM EDTA; 7 mM 2-mercaptoetyanol; 100 μg/ml BSA; and 50% glycerol;
Store at: -20°C.
Ligation: After 2-fold MspR9 I overdigestion >5% of the DNA fragments can be ligated and recut.
Non-specific hydrolisis: No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 40 u.a. of enzyme for 16 hours at 37°C.
Reagents supplied with enzyme: 10 X SE-buffer O
Methylation sensitivity: Blocked by overlapping Dcm methylation (CmCWGG): CCAGG and CCTGG.
Inactivation: 20 minutes under 80°C
MspR9 I - CC↑NGG - GGN↓CC - Restriction Endonuclease from Sibenzyme
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