Acl I - AA↑CGTT - TTGC↓AA

Source: Acinetobacter calcoaceticus

 

 

 

 

 

 

 

Recognition site:

AA↑CGTT

TTGC↓AA

 

Source: Acinetobacter calcoaceticus

 

Assayed on: Lambda DNA

 

Unit definition: One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 37°C in a total reaction volume of 50 μl.

 

Optimal SE-buffer: Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.) + BSA

 

 

Activity in SEBuffers:

SEBuffer B      0 -  10%

SEBuffer G      0 -  10%

SEBuffer O      0 -  10%

SEBuffer W      0 -  10%

SEBuffer Y           100%

SEBuffer ROSE    80%

 

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-mercaptoethanol; 200 μg/ml BSA; 0.05% Triton X-100; 50% glycerol;

 

Store at: -20°C.

 

Ligation: After 2-fold overdigestion with enzyme 90% 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 2 u.a. of enzyme for 16 hours at 37°C.

 

Reagents supplied with enzyme: 10 X SE-buffer Y, BSA

 

Methylation sensitivity: Blocked by CG methylation

 

Inactivation: 20 minutes under 65°C

 

Notes: To obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml. Do not use BSA for long incubation.

 

E012 - Acl I - 1000 units

Acl I - AA↑CGTT - TTGC↓AA - Restriction Endonuclease from Sibenzyme

 

 * availibility of sample size may be limited

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