Creative SuperCut series is developed to finish the restriction cut within 5-15mins. It is suitable to be used in vector, DNA fragment, PCR products, and DNA genome. The Creative SuperCut series has the features as follows:

• High digestion efficiency. Reactions can be finished within 5-15min.
• Use universal buffer. Easily achieves multi cut without buffer changes.
• 100% Adaptability with buffers such as FastDigest(Thermo Scientific), CutSmart(NEB), and QuickCut(Takara)
• Color buffer is attached. Simplify the loading and testing procedures in electrophoresis.
• High-fidelity enzymes

Quality Control

• Activity Assays

1μl Supercut enzymes can digest 1μg λDNA (Hind III Digest) in 15mins in optimal temperature in 20μl system.

• Ultra-long incubation test

Incubate 1 μl SuperCut enzymes and 1 μg λDNA (HindIII digest) for 3 hours at the optimal reaction temperature. No other nuclease contamination or non-specific degradation of the substrate caused by star activity is detected.Delayed digestion may cause Star activity.

• Digestion-ligation-re-digestion detection

At the optimum reaction temperature, use 1 μl SuperCut enzymes to digest the substrate and use an appropriate amount of Fast T4 DNA Ligase at 22°C to re-ligate the digested product. After the ligated product is recovered again, use the same restriction enzyme can re-cut the ligation product.

• Non-specific endonuclease activity detection

Incubate 1 μl of SuperCut enzyme and 1 μg of supercoiled plasmid DNA for 4h at the optimal reaction temperature. Use agarose gel electrophoresis to detect that the plasmid DNA, the plasmid is still in a supercoiled state.

• Blue and white spot detection

The vector containing a single lacZα gene was digested with 1 μl SuperCut enzymes. Then the vector is re-ligated and transformed into E. coli competent cells. Spread the product on the LB medium plate containing the corresponding antibiotics, IPTG and X-gal. The correctly connected product will grow blue colonies, while the wrongly connected product (that is, the DNA end cut is incomplete) will give white colonies. For SuperCut series restriction enzymes, the proportion of white colonies is less than 1%.

Samples of reaction system

• DNA SuperCut System

1. Prepare the reaction system as suggested below
2. Gently pipette or flick the tube wall to mix (do not vortex), and then centrifuge briefly to collect the wall-hanging droplets.
3. Incubate at 37°C for 15 min (plasmid), or 15-30 min (PCR product), or 30-60 min (genomic DNA);
4. Incubate at 80°C for 20 minutes to inactivate the enzyme and stop the reaction (optional).

• Double or Muti Cut system

1. The dosage of each fast endonuclease is 1 μl, and the reaction system should be appropriately expanded as needed.
2. The total volume of all fast endonucleases should not exceed 1/10 of the total reaction system.
3. If the optimum reaction temperature of the several fast endonucleases used is different, the digestion should be started with the enzyme with the lower optimum temperature first, and then the enzyme with the higher optimum temperature should be added, and the digestion reaction should be carried out at the optimum reaction temperature.