Description
PURPOSE
«ALPREP Plasmid Magnetic» Extraction kit is designed for the extraction of high-purity plasmid DNA from gram-negative bacteria using magnetic sorption method. 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
Assay principle is based on lysis of bacterial cells followed by alkali denaturation of DNA. After neutralization of alkali solution, the circular DNA restores its original structure, while genomic DNA rehybridizes randomly, forming large aggregates that precipitate together with proteins. Next, in the presence of chaotropic agents, plasmid DNA reversibly binds to the surface of magnetic particles, after which it is washed of possible contaminants and eluted in low-salt buffer. RNAse treatment allows to get rid of RNA contamination in 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.
