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OPTIZYME™ SacI, Fisher BioReagents™

Product Code. 11914782
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Quantity:
2000 U
Unit Size:
Each
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Product Code. 11914782

Brand: Fisher BioReagents BP80161

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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
Enzyme Activity in OPTIZYME buffers:
  • Buffer 1: 50 - 100%
  • Buffer 2: 20 - 50%
  • Buffer 3: 0 - 20%
  • Buffer 4: 50 - 100%
  • Buffer 5: 0 - 20%
Storage Buffer:
  • 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
  • Ligation and Re-cleavage:
  • More than 95% of the DNA fragments can be ligated and re-cut after a 50-fold over-digestion with SacI.
  • Digestion of Agarose-embedded DNA:
  • A minimum of 5 units of SacI is required for the complete digestion of 1μg of agarose-embedded lambda DNA in 16 hours.
  • Notes:
    • 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.
    Compatible Ends: Alw21I, Eco24I, SduI
    Methylation Effects:
    Dam: Never overlaps - no effect
    Dcm: Never overlaps - no effect
    CpG: May overlap - no effect
    • 5'...m5C GAGCTm5C G...3'
  • 3'... Gm5CTCGA Gm5C...5'
  • (No effect on cleavage)
  • EcoKI: Never overlaps - no effect
  • EcoBI: May overlap - no effect
    • 5'...TGm6AGCTC(N)4 TGCT...3'
  • 3'...AC TCGAG(N)4 m6ACGA...5'
  • (No effect on cleavage)

    • TRUSTED_SUSTAINABILITY

    Specifications

    Concentration 10 U/μL
    Components 10X OPTIZYME™ Ecl136II, SacI Buffer
    Incubator Temperature 37°C
    pH 7.4
    For Use With (Application) >95% of DNA fragments can be ligated and re-cut after 50-fold over-digestion with SacI
    Storage Buffer 10mM Tris HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/mL BSA, 50% Glycerol
    Quantity 2000 U
    Cut Site GAGCT.C
    Product Type SacI
    Form Liquid
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