OPTIZYME™ SacI, Fisher BioReagents™

5'...G A G C T^C...3'
3'...C^T C G A G...5'

Supplied with: 10X OPTIZYME Ecl136II, SacI Buffer
Conditions for 100% Activity:

• 1X OPTIZYME Buffer Ecl136II, SacI: 10mM Bis-Tris Propane-HCl (pH 6.5 at 37°C), 10mM MgCl₂ and 0.1mg/ml BSA
• Incubate at 37°C

• Buffer 1: 50 - 100%
• Buffer 2: 20 - 50%
• Buffer 3: 0 - 20%
• Buffer 4: 50 - 100%
• Buffer 5: 0 - 20%

• 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM DTT, 1 mM EDTA, 0.2mg/ml BSA and 50% (v/v) glycerol

• 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.

• 5'...m5C GAGCTm5C G...3'

• 5'...TGm6AGCTC(N)₄ TGCT...3'