Description
PURPOSE
«ALPREP Plasmid Column» Extraction kit is designed for the extraction of high-purity plasmid DNA from gram-negative bacteria using spin columns. Extracted plasmid DNA is free from salts, proteins, RNA, and other cellular contaminants, making it suitable for subsequent use in PCR, qPCR, restriction enzyme digestion, ligation, sample preparation for Sanger and NGS sequencing, as well as various other genetic engineering applications.
ASSAY PRINCIPLE
Principle of reagent kit is based on reversible binding of plasmid DNA to the membrane surface of a spin-column following sequential treatment with the kit’s components.
The DNA extraction and purification protocol includes:
- Cell Pelleting: Precipitation of bacterial cells.
- Resuspension: Treatment with specific solutions followed by resuspension.
- Column Incubation: Incubation in the spin column.
- Elution: Elution process followed by centrifugation to collect the purified DNA.
- RNase Treatment: Treatment with RNase to remove RNA contamination from the plasmid DNA samples.
ADVANTAGES
- Complete removal of cellular components;
- Fast and economical: extraction and purification time is 30 minutes;
- Plasmid DNA yield up to 20 µg;
- Extraction efficiency of more than 85%;
- Volume of eluted plasmid DNA 30 – 100 µl;
- Safety: chloroform and phenol are not used;
- Shelf life – 2 years.
