5'			G	A	G	C	T ↓	C		3'
3'			C ↑	T	C	G	A	G		5'

Thermo Scientific SacI restriction enzyme recognizes GAGCT^C sites and cuts best at 37°C in its own unique 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. Isoschizomers: Eco53kI, EcoICRI, Psp124BI, SstI.

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: SacI is sensitive to cytosine methylation at GAGmCTC, but not GAGCTmC and insensitive to adenine methylation at GmAGCTC. AluI methyltransferase (AGmCT) can be used to block SacI. Supercoiled plasmids may require up to 5-fold more SacI for complete digestion than linear DNA. SacI is inhibited by common clinical anticoagulants found in some preparation of anticoagulated peripheral blood and bone marrow. Levels of EDTA and ACD (citric acid-sodium citrate-dextrose) in standard sample preparation have been shown to inhibit SacI. Three times the normal concentration for heparin is required to inhibit SacI. (J. E. Coad et al., Inhibition of restriction endonucleases by common clinical anticoagulants. Anal. Biochem. 205, 368-369, 1992). For methylation sensitivity, refer to product specifications.