| 5 | G | G | C | C | N | N | N | N ↓ | N | G | G | C | C | 3 | ||||
| 3 | C | C | G | G | N ↑ | N | N | N | N | C | C | G | G | 5 |
Thermo Scientific SfiI restriction enzyme recognizes GGCCNNNN^NGGCC sites and cuts best at 50°C in G buffer. See Reaction Conditions for Restriction Enzymes for a table of enzyme activity, conditions for double digestion, and heat inactivation for this and other restriction enzymes. Note: Also available as a FastDigest enzyme for rapid DNA digestion.
Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.
Features
• Superior quality—stringent quality control and industry leading manufacturing process
• Convenient color-coded Five Buffer System
• Includes universal Tango buffer for double-digestions
• BSA premixed in reaction buffers
• Wide selection of restriction endonuclease specificities
Applications
• Molecular cloning
• Restriction site mapping
• Genotyping
• Southern blotting
• Restriction fragment length polymorphism (RFLP)
• SNP
Note: Assayed using pUC19 DNA with insert containing two SfiI sites from Adenovirus-2 DNA. Incubation at 37°C results in 10% activity. SfiI cleavage is impaired by overlapping dcm methylation. To avoid dcm methylation, use a dam-, dcm- strain, ?such as GM2163. For cleavage with SfiI, at least two copies of its recognition sequence are required. The two sites can be on either the same or different DNA molecules. (L. M. Wertzell et al., J. Mol. Biol., 248, 581-595, 1995.) Therefore, also an oligonucleotide harboring a SfiI recognition site can be supplemented. For methylation sensitivity, refer to product specifications.