Product Overview
HYF289 SacII Restriction Enzyme is a restriction endonuclease designed for DNA cleavage in molecular biology applications. It digests suitable DNA substrates in 5–15 minutes under recommended reaction conditions, with an optimum reaction temperature of 37°C.
The enzyme can be used for restriction analysis of plasmid DNA, genomic DNA, viral DNA, and PCR products containing the appropriate recognition sequences. Efficient cleavage generally requires two SacII recognition sites, so digestion may proceed more slowly when the substrate contains only one site. DNA methylation, particularly CpG methylation, and high salt concentrations may also affect cleavage efficiency.
The supplied Color Buffer contains a density reagent and red and yellow tracking dyes, allowing the digestion mixture to be loaded directly onto an agarose gel without adding a separate loading dye. Following digestion, the enzyme can be heat-inactivated at 80°C for 20 minutes, according to the recommended protocol.
Components
| Components | 50 rxns |
| Velocut SacII | 50 μL |
| 10× Buffer | 1 mL |
| 10× Color Buffer | 1 mL |
Product Features
- Rapid DNA Digestion: Digests suitable DNA substrates in 5–15 minutes under recommended reaction conditions, with an optimum reaction temperature of 37°C.
- Convenient Gel Loading: Supplied with standard reaction buffer and Color Buffer, which enables direct loading of digestion mixtures onto agarose gels without adding a separate loading dye.
- Defined Heat Inactivation: Can be heat-inactivated at 80°C for 20 minutes, helping simplify downstream workflow planning.
FAQ
Q1: What are the recommended reaction temperature and digestion time for HYF289 SacII Restriction Enzyme?
HYF289 SacII Restriction Enzyme has an optimum reaction temperature of 37°C and can digest suitable DNA substrates in 5–15 minutes under recommended reaction conditions. Actual digestion efficiency depends on the number of recognition sites, DNA methylation status, and reaction composition.
Q2: Can DNA methylation affect HYF289 SacII Restriction Enzyme activity?
Yes. Cleavage may be blocked or impaired when the substrate DNA is methylated by a CpG methylase. If digestion is incomplete, consider the methylation status of the DNA and verify that the substrate is compatible with SacII cleavage.
Q3: Why may HYF289 SacII Restriction Enzyme digest DNA containing only one recognition site more slowly?
SacII generally requires two recognition sequences for efficient cleavage. Digestion may therefore proceed more slowly when a DNA substrate contains only one SacII site. High salt concentrations may also inhibit enzyme activity, so DNA samples should be prepared in a suitable reaction-compatible buffer.
