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OPTIZYME™ SacI, Fisher BioReagents™
Brand: Fisher Bioreagents BP8016-1
Additional Details : Weight : 0.01000kg
Description
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.
Methylation Effects:
Dam: Never overlaps - no effect
Dcm: Never overlaps - no effect
CpG: May overlap - no effect
- 5'...m5C GAGCTm5C G...3'
- 5'...TGm6AGCTC(N)4 TGCT...3'
Specifications
| 10 U/μL | |
| 37°C | |
| >95% of DNA fragments can be ligated and re-cut after 50-fold over-digestion with SacI | |
| 2000 U | |
| SacI |
| 10X OPTIZYME™ Ecl136II, SacI Buffer | |
| 7.4 | |
| 10mM Tris HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/mL BSA, 50% Glycerol | |
| GAGCT.C | |
| Liquid |