The LumiPure Plasmid spin miniprep Kit is designed for rapid and efficient purification of plasmid DNA from Escherichia coli cultures using spin columns. First, cell pellet is resuspended, and then alkaline-cell lysis is performed using Lysis Solution. The lysate is mixed with Neutralization Buffer that allows selective binding of DNA to the membrane in the column at a high chaotropic agent concentration. Denatured proteins, genomic DNA and SDS are pelleted by centrifugation, and the supernatant containing the plasmid DNA is loaded onto the spin column. In the following step, a series of washings of bound DNA is performed. Special formulations of Washing Solutions allow impurities to be completely removed from the column membrane. Finally, the genomic DNA is eluted with Elution Buffer or, if necessary, with deionized water.
Samples: E. coli cells carrying plasmid DNA
Operation time: 15 minutes
Column binding capacity: 30 µg DNA
DNA typical yield: up to 30 µg depending on the plasmid copy number and the processed culture volume
DNA Purity: OD260/OD280 >1.8
The purified DNA is suitable for any subsequent application, such as PCR, restriction enzyme digestion, ligation and transformation, in vitro transcription, preparing samples for Sanger sequencing and NGS, etc.