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OPTIZYME™ NheI, Fisher BioReagents™

Product Code. 11944782
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Quantity:
500 U
Unit Size:
Each
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11944782 500 U Each
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Product Code. 11944782 Supplier Fisher BioReagents Supplier No. BP80191
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Description

5'...G^C T A G C...3'
3'...C G A T C^G...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: 100%
  • Buffer 2: 20 - 50%
  • Buffer 3: 0 - 20%
  • Buffer 4: 100%
  • Buffer 5: 0 - 20%
Storage Buffer:
  • 10mM Tris-HCl (pH 8.0 at 25°C), 50mM KCl, 1mM DTT, 0.1mM 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 NheI.
  • Digestion of Agarose-embedded DNA:
  • A minimum of 5 units of NheI is required for the complete digestion of 1μg of agarose-embedded lambda DNA in 16 hours.
  • Notes:
    • Low salt, high glycerol (>5%) concentrations, pH>8.0 or a large excess of enzyme may result in star activity.
    • NheI is inhibited by salt concentrations above 100mM.
    • Supercoiled plasmids may require up to 10-fold more NheI for complete digestion than linear DNAs (e.g. 10 units are required to cleave 1μg of pBR322 DNA).
    Compatible Ends: XmaJI, BcuI, Eco130I, XbaI
    Methylation Effects:
    Dam: Never overlaps - no effect
    Dcm: Never overlaps - no effect
    CpG: May overlap - cleavage impaired
    • 5'...GCTAGm5C G...3'
  • 3'...CGATC Gm5C...5'
  • (Cleavage is impaired)
    • 5'...m5C GCTAGm5C G...3'
  • 3'... Gm5CGATC Gm5C...5'
  • (Cleavage is blocked)
  • EcoKI: Never overlaps - no effect
  • EcoBI: Never overlaps - no effect

    • TRUSTED_SUSTAINABILITY

    Specifications

    Concentration 10 U/μL
    Components 25 to 50% Water, >50% Glycerin
    pH 7.4
    For Use With (Application) >90% of DNA fragments can be ligated and re-cut after 50-fold over-digestion with Nhe I
    Storage Buffer 10mM Tris HCl (pH 8 at 25°C), 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/mL BSA, 50% Glycerol
    Content And Storage Keep container tightly closed
    Quantity 500 U
    Cut Site G.CTAGC
    Product Type NheI
    Form Liquid
    Product Title
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