The innuPREP Plasmid Mini Kit 2.0 allows the fast and easy isolation of plasmid DNA from bacterial cultures. To gain an optimal result, protocol steps as well as the chemistry and spin filters were improved in comparison to the precursor products innuPREP Plasmid Mini Kit and innuPREP Plasmid Mini Kit Plus. The process is based on the combination of an alkaline lysis and binding of the plasmid DNA to a filter membrane after precipitation of the chromosomal DNA and bacterial proteins. Bound plasmid DNA is then washed and finally eluted by using a low-molar buffer. The isolated plasmid DNA can be used for a number of sensitive downstream applications, like transfection, cloning, PCR or sequencing. The new procedure also allows for processing of 0.5 ml to 15 ml of bacterial cultures using a table-top centrifuge (after lysis). Due to the new filter membrane up to 80 µg pDNA can be isolated from one sample. The innuPREP Plasmid Mini Kit 2.0 allows the fast and easy isolation of plasmid DNA from bacterial cultures. To gain an optimal result, protocol steps as well as the chemistry and spin filters were improved in comparison to the precursor products innuPREP Plasmid Mini Kit and innuPREP Plasmid Mini Kit Plus. The process is based on the combination of an alkaline lysis and binding of the plasmid DNA to a filter membrane after precipitation of the chromosomal DNA and bacterial proteins. Bound plasmid DNA is then washed and finally eluted by using a low-molar buffer. The isolated plasmid DNA can be used for a number of sensitive downstream applications, like transfection, cloning, PCR or sequencing. The new procedure also allows for processing of 0.5 ml to 15 ml of bacterial cultures using a table-top centrifuge (after lysis). Due to the new filter membrane up to 80 µg pDNA can be isolated from one sample.