Kits
NucleoBond® Xtra Midi, typical yield ≥ 250 µg plasmid DNA
NucleoBond® Xtra Maxi, typical yield ≥ 1000 µg plasmid DNA
NucleoBond® Xtra Midi/Maxi Plus, NucleoBond® Finalizer/Finalizer Large included to speed up DNA precipitation. Saving of time!
Fig. 1.
Features
the new generation of anion exchangers
up to 60% time saving
up to 100% increased yields of transfection-grade plasmid DNA
highest purity of plasmid DNA due to anion-exchange technology
MACHEREY-NAGEL has developed NucleoBond® Xtra, a new generation of anion exchangers based on the patented NucleoBond® technology.
With NucleoBond® Xtra typical yields of ≥ 250 µg (Midi) or ≥1000 µg (Maxi) of ultra-pure plasmid DNA can be obtained in about half of the time compared to other Midi and Maxi kits based on anion-exchange chromatography.
Fig. 1. (Click to enlarge)
NucleoBond® Xtra Midi and Maxi kits contain enlarged columns which leads to lower silica resin beds. This in turn enables a faster flow of lysate and buffers through the columns.
Specially designed column filters are included for convenient and time-saving clarification of bacterial lysates. The column filters are supplied directly inserted in the NucleoBond® Xtra columns and allow parallel clarification of bacterial lysate and loading onto the column. Their large, structured surface leads to high filter flow rates and minimized risk of clogging.
The improved silica material with greater DNA binding capacity is based on the proven NucleoBond® anion-exchanger group MAE (methylaminoethanol). Optimized buffer compositions additionally lead to improved alkaline lysis and increased column flow rates.
Procedure
Bacteria are harvested from an overnight culture and lysed by an optimized alkaline lysis procedure. The bacterial lysate is cleared and loaded onto the equilibrated column in one step! The 1st washing step is performed with inserted column filter in order to obtain maximum recovery of DNA. After subsequent washing, the purified plasmid DNA is eluted in a high-salt buffer and precipitated with isopropanol. Precipitation can be performed by centrifugation (NucleoBond® Xtra Midi or Maxi) or speeded-up by the use of the NucleoBond® Finalizer/Finalizer Large included in the NucleoBond® Xtra Midi Plus/Maxi Plus kits.
Fig. 2. (Click to enlarge)
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