QIAGEN Plasmid Plus 96 Kits

QIAGEN Plasmid Plus 96 Kits provide a novel method for plasmid preparation in 96-well format. Samples can be conveniently processed using the QIAvac 96 and/or a centrifuge or automated on the BioRobot Universal System. Proprietary binding chemistry allows up to 50 μg of transfection-grade plasmid DNA per well to be obtained from up to 5 ml of an E. coli culture. Endotoxin levels are reduced to levels below 1 EU/µg DNA due to a dedicated endotoxin reduction buffer.
For optimal results it is recommended to use this product together with QIAvac 96.

Consistently high yields of ready-to-use plasmid DNA

QIAGEN Plasmid Plus technology delivers the same performance and quality as anion-exchange technology. High yields of up to 50 μg of transfection-grade plasmid DNA can be obtained (see figure " High yields of plasmid DNA").

Very low endotoxin levels

Kits include an endotoxin reduction buffer that effectively reduces bacterial endotoxins to levels typically lower than 1 EU/μg DNA, providing plasmid DNA quality which is suitable for transfection into sensitive cell lines (see figures " Very low endotoxin levels" and " Highly efficient transfection into a sensitive cell line").

QIAGEN Plasmid Plus 96 Kits are based on QIAGEN Plasmid Plus technology, a novel method for plasmid preparation. The procedure is based on a nonchaotropic binding chemistry that ensures selectivity and DNA quality that is comparable to anion exchange preps. Following lysate clearing, a simple bind-wash-elute procedure results in highly concentrated plasmid DNA, ready for direct use in subsequent applications. The unique kit chemistry and design of the Plasmid Plus 96 plates allow high binding capacities.

This novel method for plasmid preparation in 96-well format provides highly pure, transfection-grade plasmid DNA using a simple bind-wash-elute procedure (see flowchart " QIAGEN PlasmidPlus 96 procedure").

The QIAGEN Plasmid Plus 96 Miniprep protocol starts with a modified alkaline lysis procedure. After neutralization, lysates are cleared using a TurboFilter 96 plate. A nonchaotropic binding buffer is added to the cleared lysate to optimize plasmid DNA binding to the membrane of the Plasmid Plus 96 plate. The unique binding buffer provides very specific binding conditions leading to DNA quality that is comparable to anion-exchange preps. A wash step with an endotoxin reduction buffer results in transfection-grade plasmid DNA suited for transfection into most cell lines (see figure " Highly efficient transfection into a sensitive cell lines"). The proprietary binding chemistry allows purification of up to 50 μg plasmid DNA per well from a 5 ml E. coli culture (see figure " High yields of plasmid DNA").

Convenient and flexible processing options

The procedure may be fully automated using the QIAGEN Plasmid Plus 96 BioRobot Kit on the BioRobot Universal System. Alternatively, the QIAvac 96 is used for lysate clearing using the TurboFilter 96 plate, and DNA binding and wash steps are performed using the Plasmid Plus 96 plate. Drying and elution steps may either be performed on the QIAvac 96 or using a compatible centrifuge. In addition, the complete procedure can be performed using a compatible centrifuge.

Plasmid DNA purified using QIAGEN Plasmid Plus 96 Kits is highly suitable for a wide variety of applications, including:

• Transfection, including into sensitive cell lines
• Preparation of short hairpin vectors (sh-vectors)
• Enzymatic modifications
• Cloning
• Restriction digestion