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OPTIZYME™ PfoI, Fisher BioReagents™

Product Code. 11924832
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Quantity:
200 U
Unit Size:
Each
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Product Code. 11924832 Supplier Fisher BioReagents Supplier No. BP80771
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Description

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

Supplied with: 10X OPTIZYME Buffer 4
Conditions for 100% Activity:

  • 1X OPTIZYME Buffer 4: 33mM Tris-acetate (pH 7.9 at 37°C), 10mM Mg-acetate, 66mM K-acetate and 0.1mg/mL BSA
  • Incubate at 37°C
Enzyme Activity in OPTIZYME buffers:
  • Buffer 1: 0 - 20%
  • Buffer 2: 20 - 50%
  • Buffer 3: 50 - 100%
  • Buffer 4: 100%
  • Buffer 5: 0 - 20%
Storage Buffer:
  • 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM DTT, 1mM 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 50-fold over-digestion with PfoI.
  • Digestion of Agarose-embedded DNA:
  • A minimum of 10 units of PfoI is required for the complete digestion of 1μg of agarose-embedded lambda DNA in 16 hours.
  • Note:
    • PfoI cleavage is impaired by overlapping dam methylation and blocked by overlapping dcm methylation. To avoid dam or dcm methylation, use a dam-, dcm- strain.
  • Compatible Ends: EcoRII, SexAI, BssKI
  • Methylation Effects:
  • Dam: May overlap - cleavage impaired
    • 5'...TCCNGGm6A TC...3'
  • 3'...AGGNCC Tm6AG...5'
  • (Cleavage is impaired)
  • Dcm: May overlap - blocked
    • 5'...TCm5CA GGA...3'
  • 3'...AG GTm5CCT...5'
  • (Cleavage is blocked)
  • CpG: May overlap - blocked
    • 5'...TCm5C GGGA...3'
  • 3'...AG Gm5CCCT...5'
  • (Cleavage is blocked)
  • EcoKI: Never overlaps - no effect
  • EcoBI: Never overlaps - no effect

    • TRUSTED_SUSTAINABILITY

    Specifications

    Concentration 10 U/μL
    Components >50% Glycerin, 25 to 50% Water
    Incubator Temperature 37°C
    pH 7.4
    For Use With (Application) >90% of DNA fragments can be ligated and re-cut after 10-fold over-digestion with Pfo I
    Storage Buffer 10mM Tris HCl (pH 7.4 at 25°C), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/mL BSA, 50% Glycerol
    Content And Storage Keep container tightly closed
    Quantity 200 U
    Cut Site T.CCNGGA
    Product Type PfoI
    Form Liquid
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