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OPTIZYME™ DpnI, Fisher BioReagents™

Product Code. 11904852
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Quantity:
500 U
Unit Size:
Each
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Product Code. 11904852

Brand: Fisher BioReagents BP80091

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Description

5'...G m6A^T C...3'
3'...C T^m6A 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: 100%
  • Buffer 3: 50 - 100%
  • Buffer 4: 100%
  • Buffer 5: 50 - 100%
Storage Buffer:
  • 10mM Tris-HCl (pH 7.4 at 25°C), 400mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 50% (v/v) glycero
l
  • Ligation and Re-cleavage:
  • More than 70% of the pBR322 DNA fragments can be ligated and more than 95% of these can be re-cut after 50-fold over-digestion with DpnI.
  • Notes:
    • DpnI requires the presence of N6-methyladenine within the recognition sequence to cleave DNA.
    • DpnI will only cleave fully-adenomethylated dam sites. Hemi-adenomethylated dam sites DpnI cleaves 60X more slowly.
    • DpnI, Bsp143I and MboI all recognize the same sequence but have different methylation sensitivities and cleavage sites.

    Methylation Effects:
    Dam: Does not cut dam- DNA
    • Gm6ATC
  • (Cleaves only Dam methylated DNA)
  • Dcm: Never overlaps - no effect
  • CpG: May overlap - no effect
    • 5'...Gm6A Tm5C G...3'
  • 3'...C Tm6A Gm5C...5'
  • (Cleaves only Dam methylated DNA)
  • EcoKI: Never overlaps - no effect
  • EcoBI: May overlap - effect not determined

    • TRUSTED_SUSTAINABILITY

    Specifications

    Concentration 10 U/μL
    Components 10X OPTIZYME™ Buffer 4
    Incubator Temperature 37°C
    pH 7.4
    For Use With (Application) >70% of pBR322 DNA fragments can be ligated and >95% of these can be re-cut after 50-fold over-digestion with DpnI
    Storage Buffer 10mM Tris HCl (pH 7.4 at 25°C), 400mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/mL BSA, 50% Glycerol
    Content And Storage Keep container tightly closed
    Quantity 500 U
    Cut Site GA.TC
    Product Type DpnI
    Form Liquid
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