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.
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