Principle

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.
NucleoBond® Xtra Midi and Maxi kits contain enlarged columns which leads to lower silica resin beds. This in turn enables 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 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 (patented technology). Optimized buffer compositions additionally lead to improved alkaline lysis and increased column flow rates.

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 (plasmid DNA binds to the anion-exchange resin). The first washing step using equilibration buffer is performed with inserted column filter to wash out residual lysate from the filter and 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 by using the NucleoBond® Finalizer / Finalizer Large included in the NucleoBond® Xtra Midi Plus/Maxi Plus kits to reduce the total prep time to about 30 minutes (Midi Plus) or 35 minutes (Maxi Plus).
 

Features

The new generation of anion exchanger – ultra-fast, transfection-grade plasmid DNA with high yield

• Optimized column design – Midi prep in approximately 30 min, Maxi prep in 35 min
• Column filter included – high filter flow rates, parallel lysate clearing, and loading onto the column
• Improved silica material – typical yields of 400 μg (Midi) and 1 000 μg (Maxi)
• Proven anion-exchange technology – transfection-grade plasmid DNA
• NucleoBond® Xtra Plus kits contain NucleoBond® Finalizers to speed up DNA precipitation

Technical specifications