NotI-HF Restriction Enzyme

NotI-HF restriction enzyme recognizes GC^GGCCGC site and cuts best at 37°C in 5-15 minutes using Reaction Buffer provided. NotI-HF has the same specificity as native enzyme, but has been engineered for significantly reduced star activity.

The enzyme is available in 500 and 2,500 unit sizes at a concentration of 20 U/µL. The enzyme is supplied with a 10× Reaction Buffer.

Applications

• Molecular cloning.
• Restriction site mapping.
• Genotyping.
• Southern Blot.

Product Specifications

• Storage Buffer: 10 mM Tris-HCl (pH 7.4), 50 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 0.2 mg/mL BSA, and 50% (v/v) glycerol.
• Unit Definition: One unit is defined as the amount of enzyme required to digest 1 μg of pBC4 DNA in 1 hour at 37°C in a total reaction volume of 50 μl.

Restriction Digest Protocol

1.  Assemble the following reaction at room temperature:

          10× Reaction Buffer                        5 µL
          DNA                                                1 µg
          NotI-HF                                           0.5-1 µL
          Nuclease-free water                       to 50 µL
          Total volume                                   50 µL

2.  Mix gently and spin down for a few seconds.

3.  Incubate at 37 °C for 1 h.

4.  Stop the reaction by heating at 70°C for 10 min.