SweFast Cut NheI, 100 T (for Restriction Endonucleases)

Product Information

Product Description

SweFast Cut NheI is expressed and purified from E. coli strain carries the recombinant NheI gene cloned from Neisseria mucosa heidelbergensis. SweFast Cut NheI is a fast restriction endonuclease with the recognition site at G^CTAGC. The reaction is conducted for 5-15 minutes at 37, and heat-inactivated at 80°C for 20 minutes. This enzyme is not sensitive to dam methylation, dcm methylation, but sensitive to CpG methylation. SweFast Cut NheI is suitable for fast cutting plasmid DNA, PCR products, genomic DNA, etc.

Unit Definition: One unit defined as the amount of enzyme required to digest 1 µg of λDNA (HindIII digest) in a total reaction volume of 20 µL at 37°C within 15 minutes.

Reaction Condition: 1×SweFast Cut Buffer, Incubate at 37°C.

Storage Solution: 10 mM Tris-HCl, 250 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 200 µg/ml BSA, 50% Glycerol

0.15% Triton X-100, pH 7.4 @ 25°C.

10×SweFast Cut Buffer: 500 mM Potassium Acetate, 200 mM Tris Acetate, 100 mM Magnesium Acetate, 1000 µg/ml BSA, pH 7.9 @ 25°C.

Storage and Shipping Conditions

Shipped with dry ice and stored at -20 with a validity period of 2 years.

Product Content

Assay Protocol

1. Recommended System for Single Digestion

1) Prepare the reaction system on the ice by adding samples as suggested in the table, a: This system is suitable for enzymatic digestion of purified PCR products;

2) Mix the 10×SweFast Cut Buffer before use;

3) Gently tap or flick the tube to mix (do not vortex), and then centrifuge instantaneously;

4) Incubation at 37 for 5-15 minutes (plasmid), or 15-30 minutes (PCR products), or 30-60 minutes (genomic DNA), Different DNA structure will lead to different enzyme digestion effect, and the reaction time can be adjusted according to the effect of enzyme digestion.

5) If the enzyme digestion is incomplete or DNA exceeds 2 μg, the amount of restriction endonuclease can be appropriately increased, but the total amount of restriction endonuclease should not exceed 10% of the total reaction system.

6) Incubation at 80 for 20 minutes can deactivate the enzyme and stop the reaction (optional). For enzymes that are not heat-inactivated, we recommend column purification or recovery after agarose gel electrophoresis.

7) If the total reaction volume is more than 20 μL, the incubation time should be appropriately increased, and the water bath, metal bath or sand bath should be used as far as possible.

2. Recommended System for Double Digestion

1) Add 1 μL of each fast restriction endonuclease, and the reaction system should be appropriately expanded as required;

2) The total volume of all fast restriction endonuclease should not exceed 10% of the total reaction system, and the concentration of glycerol in the reaction system should be <5% to avoid asterisk activity. For example, the total enzyme amount in the 50 μL reaction system does not exceed 5 μL.

3) If the optimal reaction temperature of several fast restriction endonuclease used is different, the enzyme with the lowest optimal temperature should be added first, and then add the enzyme with the highest optimal temperature after inactivation of the first enzyme, and the enzyme digestion reaction should be carried out at its optimal reaction temperature.

4) Incubation at the recommended reaction temperature and time, and overnight incubation is not recommended for the double enzyme digestion reaction.

Note

1. There was no asterisk activity after 3 h incubation, and asterisk activity may occur after delayed digestion.

2. For your safety and health, please wear a lab coat and disposable gloves. 

For Research Use Only!

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