Genopure Plasmid Midi Kit
The Genopure Plasmid Midi Kit is used to prepare purified plasmid DNA in medium quantities known as “midi preps”. Using a modified alkaline lysis method, highly purified plasmid DNA from E. coli (free of RNA contamination) is generated. The kit is designed for the isolation of up to 100 µg of plasmid DNA from bacterial culture. Depending on the number of plasmids, use a 5-ml to 30-ml (high-copy number) or 10-ml to 100-ml (low-…
The Genopure Plasmid Midi Kit is used to prepare purified plasmid DNA in medium quantities known as “midi preps”. Using a modified alkaline lysis method, highly purified plasmid DNA from E. coli (free of RNA contamination) is generated.
The kit is designed for the isolation of up to 100 µg of plasmid DNA from bacterial culture. Depending on the number of plasmids, use a 5-ml to 30-ml (high-copy number) or 10-ml to 100-ml (low-copy number) bacterial suspension.
The purified plasmid DNA from E. coli is suitable for all molecular biology applications (e.g., transfection, PCR, restriction enzyme digestion/Southern blotting, sequencing, and cloning).
Principle
This kit relies on alkaline lysis to free the plasmid DNA from bacterial cells, leaving behind E. coli “chromosomal” DNA trapped in the cell wall debris. After the solution is cleared of cell debris and “chromosomal” DNA, the supernatant is retained and passed through a pre-equilibrated column. The DNA is bound to an ion-exchange silica material packed in a gravity flow column. Low salt concentrations are needed for the binding of DNA to the macroporous anion exchanger.
Cellular impurities are removed by increased salt concentrations of the wash buffer, then the plasmid DNA is eluted. A precipitation step is added to remove salt and concentrate the eluate.
