Product Overview
Restriction digestion is one of the most frequent steps in cloning and DNA analysis, and it is often a bottleneck. Standard enzymes can need an hour or more, and combining enzymes often means switching buffers or digesting in sequence. Velocut AgeI belongs to the Velocut series of engineered restriction enzymes designed for fast DNA digestion. AgeI recognizes the sequence A^CCGGT, and Velocut AgeI digests DNA in 5 to 15 minutes. The enzyme shows superior activity in both the universal buffer and the Color Buffer, so any combination of Velocut restriction enzymes can work simultaneously in one reaction tube, with no need for sequential digestions. Velocut enzymes have passed multiple strict quality controls and can be used to digest plasmid, genomic, and viral DNA as well as PCR products. The Color Buffer contains a density reagent together with red and yellow tracking dyes, so the reaction mixture can be loaded directly onto a gel without adding loading dye. In a 1% agarose gel, the red dye migrates with 2.5 kb double-stranded DNA fragments and the yellow dye migrates with 10 bp double-stranded DNA fragments, which gives a visual reference for the run. The following isoschizomers recognize the same sequence: AsiGI, CspAI, BshTI, and PinAI. Isoschizomers may have different methylation sensitivities, so check this when replacing one enzyme with another in an existing protocol.
Components
| Components | 25 rxns |
| Velocut AgeI | 25 μL |
| 10× Buffer | 1 mL |
| 10× Color Buffer | 1 mL |
Product Features
- Fast digestion in 5 to 15 minutes: The engineered enzyme digests DNA in 5 to 15 minutes, which shortens the digestion step in cloning and analysis workflows from an hour or more to minutes.
- Multiple enzymes in one tube with the universal buffer: Superior activity in the universal buffer lets any combination of Velocut restriction enzymes work simultaneously in one reaction tube, which removes the need for buffer changes or sequential digestions.
- Color Buffer for direct gel loading: The Color Buffer includes a density reagent and red and yellow tracking dyes, so reactions go straight onto the gel. The red dye runs with 2.5 kb fragments and the yellow dye with 10 bp fragments in a 1% agarose gel.
FAQ
Q1: What is the Color Buffer, and can I load the reaction directly on a gel?
Yes. The Color Buffer contains a density reagent and red and yellow tracking dyes, so the finished reaction can be loaded directly on a gel. In a 1% agarose gel, the red dye migrates with 2.5 kb double-stranded DNA fragments and the yellow dye migrates with 10 bp double-stranded DNA fragments. Velocut enzymes work in both the universal buffer and the Color Buffer.
Q2: Can I combine Velocut AgeI with other restriction enzymes in one reaction?
Yes. All Velocut enzymes show superior activity in the universal buffer and Color Buffer, so any combination of Velocut restriction enzymes can work simultaneously in one reaction tube, and sequential digestions are not needed. Digestion takes 5 to 15 minutes. Please confirm that the recognition sites of the enzymes you combine do not conflict in your DNA.
Q3: Which DNA types can it digest, and which isoschizomers are there?
Velocut enzymes can digest plasmid, genomic, and viral DNA as well as PCR products. Isoschizomers of AgeI include AsiGI, CspAI, BshTI, and PinAI. Isoschizomers may have different methylation sensitivities, so check this when you substitute one for another in an existing protocol.
