NucleoBond Xtra plasmid kits - fast, clean, and reliable

An ever-growing range of biochemical applications requires medium to large amounts of plasmid DNA free of contamination with salts and residual bacterial components. Using NucleoBond Xtra Maxi, yields of up to 1000 μg of ultrapure plasmid DNA can be obtained based on reliable and well-established anion exchange chromatography. NucleoBond Xtra Maxi kits contain enlarged columns, which lead to lower silica resin beds. This in turn enables the 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 following features make NucleoBond Xtra Midi ideal for plasmid isolation:

• Proven anion-exchange technology for highest yields and quality,

• Optimized column design for minimum risk of clogging,

• Accelerated procedure with NucleoSpin Finalizer for a complete prep in approximately 30 min.

NucleoBond Xtra Midi procedure

Transformed (plasmid-containing) bacterial cells are lysed by an optimized set of buffers. After equilibration of the NucleoBond Xtra Column and NucleoBond Xtra Column Filter, the entire lysate is loaded by gravity flow and simultaneously cleared by the NucleoBond Xtra Column Filter. Plasmid DNA is bound to the NucleoBond Xtra Silica Resin. After efficient washing steps, the plasmid DNA is eluted and desalted, precipitated, and easily dissolved in any suitable buffer (e.g., low-salt buffer or water) for further use. Precipitation can be performed by centrifugation using the NucleoSpin Finisher to reduce the total prep time to about 30 minutes or via NucleoMag Desalting Beads.

NucleoBond Xtra anion exchange resin

NucleoBond Xtra is a patented silica-based anion exchange resin, developed by MACHEREY-NAGEL. It consists of hydrophilic, macroporous silica beads functionalist with MAE (methyl-amino-ethanol). The dense coating of this functional group provides a high overall positive charge density under acidic pH conditions, which enables the negatively charged phosphate backbone of plasmid DNA to bind with high specificity. The purified plasmid DNA is suitable for use in the most demanding molecular biology applications, including transfection, in vitro transcription, and sequencing.

Proven high yield and short preparation time

Higher yield in comparison to QIAGENs data on www.qiagen.com

Comparison: A) Kits including desalting tool, B) Kits without desalting tool

Shorter preparation time in comparison to QIAGENs data on www.qiagen.com

Comparison: A) Kits including desalting tool, B) Kits without desalting tool

LyseControl - for highest yields

LyseControl is a blue reagent premixed for your convenience with Lysis Buffer LYS. Upon addition of lysis buffer to the resuspended cells, the cell suspension turns blue (Figure 2A). The blue solution is gently inverted 5 times and incubated for 5 min to avoid contamination with genomic DNA. Upon addition of Buffer to neutralize the lysate, LyseControl turns colorless (Figure 2C). Traces of blue LyseControl (Figure 2B) indicate insufficient mixing. This step removes SDS, renatures plasmid DNA, and precipitates proteins and genomic DNA. Hence, it prevents filter clogging, maximizes plasmid DNA yield, and minimizes contamination.

Selected citations

Gieselmann et al. Effective high-throughput isolation of fully human antibodies targeting infectious pathogens. Nature Protocols 16, 3639-3671(2021).

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HiSpeed (QIAGEN GmbH, 40724 Hilden, DE)