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2
Product description
2.1
Basic principle
With the NucleoSpin
resuspended (Buffer A1) and plasmid DNA is liberated from the E. coli host cells by
SDS / alkaline lysis (Buffer A2). Buffer A3 neutralizes the resulting lysate and creates
appropriate conditions for binding of plasmid DNA to the silica membrane of the
NucleoSpin
Plasmid EasyPure Column. Precipitated protein, genomic DNA, and
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cell debris are then pelleted by a centrifugation step. The supernatant is loaded onto a
NucleoSpin
Plasmid EasyPure Column.
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With the NucleoSpin
nucleases, and soluble macromolecular cellular components are removed by only a
single washing step with Buffer AQ. Pure plasmid DNA is finally eluted under low ionic
strength conditions with slightly alkaline Buffer AE (5 mM Tris/HCl, pH 8.5).
2.2 Kit specifications
• The NucleoSpin
scale preparation of highly pure plasmid DNA (mini preps).
• The NucleoSpin
treated silica membrane which allows speeding up the procedure by a combined
washing and drying step. No additional steps are necessary if nuclease rich
host strains are used. The number of washing and drying steps is reduced from
3 to only 1!
• LyseControl: The Lysis Buffer A2 contains a blue pH indicator to ensure
complete neutralization for maximum yield.
• The purified plasmid DNA is suitable for applications like automated fluorescent
DNA sequencing, PCR, or any kind of enzymatic manipulation.
* For preparation of working solutions and storage conditions see section 3.
** Composition of Elution Buffer AE: 5 mM Tris/HCl, pH 8.5
6
Plasmid DNA purification
Plasmid EasyPure method, the pelleted bacteria are
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Plasmid EasyPure kit contaminations like salts, metabolites,
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Plasmid EasyPure kits are designed for the rapid, small-
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Plasmid EasyPure Column features a new specially
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MACHEREY-NAGEL – 08 / 2 013, Rev. 01
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