The NucleoSpin Plasmid miniprep kit is designed for the rapid (25 minutes), high-yield (up to 45 µg), small-scale preparation of high-purity plasmid DNA. These miniprep columns contain a unique silica membrane that binds up to 50 µg of plasmid DNA. The purified DNA is suitable for transfection, cloning, sequencing, PCR, transformation, and restriction analysis.

The miniprep kit allows purification of up to 25 μg of plasmid DNA per preparation, from 1–5 ml of saturated E. coli culture. You can also use this kit to purify up to 45 μg of plasmid DNA from 5–10 ml E. coli culture. In order to obtain optimal results from 5–10 ml of E. coli culture, increase your buffer volumes using the NucleoSpin Plasmid Buffer Set.

Using the NucleoSpin Plasmid miniprep kit

Harvested bacteria are resuspended and processed by SDS/alkaline lysis. High-salt buffer is added to neutralize the lysate and to create appropriate conditions for DNA binding to the silica membrane. After centrifugation, the clear supernatant is loaded onto a NucleoSpin Plasmid miniprep spin column.

Contaminants like salts and macromolecular cellular components are removed by simple washing with ethanolic Buffer A4. Additional washing with Buffer AW is recommended for the following demands:

• Complete removal of high levels of endonucleases (e.g., from wild-type E. coli strains), with Buffer AW prewarmed to 50°C
• Processing 5–10 ml of bacterial culture
• Long read length in DNA sequencing
• Better performance of critical enzymatic reactions

Highly pure plasmid miniprep DNA is finally eluted in 50 μl of slightly alkaline Buffer AE (5 mM Tris/HCl, pH 8.5). Alternatively, water (pH 8.0–8.5) can be used.