Isolate up to 200 µg transfection grade, low-endotoxin plasmid DNA (<1EU/µg) from 15-50 mL bacterial cultures using syringe filters for lysate clearance and midi spin columns
Overview
The E.Z.N.A.® Endo-Free Plasmid Midi Kit provides an efficient method for the large-scale isolation of low-endotoxin plasmid DNA. The system utilizes Lysate Clearance Filter Syringes instead of centrifugation to rapidly clear bacterial lysates post-alkaline lysis. Following lysate clearance, endotoxins are removed, and the protocol follows a simple “bind-wash-elute” procedure to deliver high-quality plasmid DNA. The yields vary according to plasmid copy number, E. coli strain, and growth conditions. 15-50 mL of bacterial cultures in LB medium typically produces up to 200 µg of high copy number plasmid DNA. Up to 100 mL of culture may be processed when working with low copy number plasmid. The system uses centrifugation or vacuum technology for plasmid purification and eliminates the time-consuming gravity-flow columns that require alcohol precipitation. Purified plasmid DNA is suitable for sensitive downstream applications such as transfection of mammalian cells, cloning, automated fluorescent DNA sequencing (typical reads exceed 800 bp), next-generation sequencing, restriction enzyme digestion, ligation, PCR, in vitro transcription, transformation, etc.
Rapid – Purification of plasmid DNA in < 45 min
Safe – No phenol/chloroform extractions
Versatile – Spin and vacuum formats available
High-quality – Transfection grade, low-endotoxin plasmid DNA (< 1 EU/µg)
Specifications
Not for use in diagnostic procedures.