BeyoFast™ KpnI

The high-quality BeyoFast™ restriction enzymes are genetically engineered for fast DNA digestion in 5-15 minutes with one buffer only. They are suitable for quick digestion of plasmid DNA, PCR products, genomic DNA, etc.

The BeyoFast™ restriction enzymes have the following advantages: (1) Digestion can be completed in 5-15 minutes. (2) All enzymes share one CutEZ™ Buffer, which simplifies the multiple enzyme digestion significantly. (3) To address the issue of varying enzyme activity in CutEZ™ Buffer, the concentration of different enzymes has been adjusted, allowing for a uniform addition of 1μl enzyme per 20μl reaction for digestion. (4) Many modifying enzymes, such as Alkaline Phosphatase, Antarctic Phosphatase, T4 DNA Ligase, T4 Polynucleotide Kinase, and T4 PNK (3′ phosphatase minus), have full activities in CutEZ™ Buffer, enabling simultaneous digestion-ligation or digestion-modification-ligation in one tube.  (5) These enzymes have good activity redundancy, making it easy to handle excessive substrates or substrates that are difficult to digest.

Enzyme activity assay: At the optimal reaction temperature, 1μl of BeyoFast™ KpnI can completely digest 1μg of λDNA (HindIII digest) in a 20μl reaction within 15 minutes.

Extended digestion assay: After digesting 1μg of λDNA (HindIII digest) with 1μl of BeyoFast™ KpnI for 3 hours at the optimal reaction temperature, no nuclease activity or non-specific cleavage caused by star activity was observed. However, extended digestion may result in star activity of restriction enzymes.

Digestion-ligation-redigestion assay: At the optimal reaction temperature, the digested substrate by BeyoFast™ KpnI can be religated with the T4 DNA Ligase at 22℃. After recovering the ligation products, they can be redigested using the same restriction enzyme.

Non-specific endonuclease activity detected: At the optimal reaction temperature, incubating 1μl of BeyoFast™ KpnI with 1μg of supercoiled plasmid DNA for 4h did not change the conformation of the plasmid DNA, as observed by agarose gel electrophoresis.

Blue-white screening: Digest 1μl of vector containing a single lacZα gene with the BeyoFast™ KpnI, followed by re-ligation and transformation into competent Escherichia coli cells. The transformants are selected on LB agar plates containing appropriate antibiotics, IPTG, and X-gal. Clones with correctly ligated products will grow as blue colonies, while those with incorrect products are white. For the BeyoFast™ series of restriction enzymes, the percentage of white colonies should be less than 1%.

The basic information of this product is as follows:

BeyoFast™ KpnI activity in different reaction buffers (buffer compatibility) is as follows:

Note: The assay results are obtained in standard reactions of BeyoFastTM restriction enzymes.

Effects of methylations on BeyoFast™ ApaLI activity are as follows:

Store at -20℃, stable for at least two years.

The restriction enzyme should be kept on ice during use, and stored at -20℃ immediately after use.

For ultrapure water without nuclease, we recommend the BeyoPure™ Ultrapure Water (DNase/RNase-free, Sterile).

If the expected enzyme digestion site cannot be cut, please check for potential methylation interference.

Isoschizomers may have different sensitivities to different methylations. When encountering potential methylation interference issues, a different restriction enzyme can be attempted.

This product is for R&D only. Not for drug, household, or other uses.

For your safety and health, please wear a lab coat and disposable gloves during the operation.