For ultra-fast small scale preparation of up to 35 µg of high-purity plasmid DNA

• LyseControl for visualization of completed alkaline lysis
• Liquid RNase A – easy handling without dissolving
• One combined washing and drying step for an ultra-fast procedure – 6 Mini preps in 14 min
  

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Procedure

The NucleoSpin® Plasmid EasyPure kit combines customer needs for highest plasmid yield with a very fast procedure.

This columns offer a binding capacity of 35 µg plasmid DNA and allow reliable yields even from higher culture volumes. The very short procedure is achieved by a combined washing and drying step. Therefore, 6 preparations can be done within 14 min.

The procedure is simple and starts with an alkaline lysis of a bacterial culture. The blue LyseControl premixed in the Lysis Buffer visualizes a completed neutralization and a finished alkaline lysis. The lysate is cleared by centrifugation and the supernatant is loaded onto the NucleoSpin® Plasmid EasyPure Column. Contaminations like salts, metabolites, nucleases, and soluble macromolecular cellular components are removed by a single washing and drying step.

Applications

• High-copy plasmids from E. coli
• Low-copy plasmids, P1 constructs, or cosmids (increased buffer volumes required)
• Plasmid DNA from Gram-positive bacteria and plasmid DNA clean-up from reaction mixtures
• Typical downstream applications: cloning, sequencing, PCR, transformation, restriction analysis

Features

Application data

High plasmid yield even with increasing culture volumes
Plasmid DNA (pcDNA3.1) was isolated from 1 mL, 2 mL, 3 mL, and 4 mL culture volumes (E. coli strain DH5α) using NucleoSpin® Plasmid EasyPure and competitor kits. The isolation was performed according to the standard procedure from each manufacturer.

Figure A: The isolated DNA was quantified by UV/VIS spectroscopy. Indicated yields are mean values of three isolations.

Figure B: Plasmid DNA was eluted in 50 µL. 2.5 µL from each eluate were analyzed on a 1 % TAE agarose gel.

Total preparation time of 6 Mini preps with NucleoSpin® Plasmid EasyPure (MN) in comparison to competitor kits

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