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

Thermo Scientific SacI restriction enzyme recognizes the GAGCT^C site and cleaves optimally at 37°C in its unique buffer. For a table of enzymatic activities, double digestion conditions, and heat inactivation of this and other restriction enzymes, see Restrictions Reaction conditions for sexual endonuclease. Note: FastDigest enzyme is also available for rapid DNA digestion. Isoschiases: Eco53kI, EcoICRI, Psp124BI, SstI.

Thermo Scientific Conventional Restriction Endonucleases are a large collection of high quality restriction enzymes optimized to function in one buffer of a five-buffer system. In addition, a universal Tango buffer is provided to facilitate double enzyme digestion. All enzymes demonstrated 100% activity under recommended buffers and reaction conditions. To ensure consistent performance, Thermo Scientific Restriction Enzyme Reaction Buffers contain premixed BSA, which enhances the stability of multiple enzymes and binds contaminants that may be present in DNA preparations.

Features

• Superior quality—strict quality control and industry-leading production processes
• Convenient color-coded five-buffer system
•Includes universal Tango buffer for double digestion
•Pre-mixed BSA in reaction buffer
•Multiple restriction endonuclease specificities available

< b>Applications

•Molecular cloning
•Restriction site mapping
•Genotyping
•Southern blot
•Restriction fragment length polymorphism ( RFLP)
•SNP

Remarks:SacI is sensitive to cytosine methylation at GAGmCTC, but is insensitive to cytosine methylation at GAGCTmC, and is sensitive to GmAGCTC Insensitive to adenine methylation. AluI methyltransferase (AGmCT) can be used to block SacI. For complete digestion, supercoiled plasmids may require up to 5 times the amount of SacI as linear DNA. SacI is inhibited by clinically common anticoagulants found in certain anticoagulated peripheral blood and bone marrow preparations. Studies have shown that levels of EDTA and ACD (citric acid-sodium citrate-glucose) in standard sample preparations have an inhibitory effect on SacI. The concentration of heparin required to inhibit SacI is three times the normal concentration. (JE Coad et al., Inhibition of restriction endonucleases by common clinical anticoagulants. Anal. Biochem. 205, 368-369, 1992). See product specifications for methylation sensitivity.