The E.Z.N.A.® FastFilter Plasmid Midi Kit combines the convenience of spin columns with the speed of syringe filters to ease large-scale plasmid preparations. The system utilizes Lysate Clearance Filter Syringes instead of centrifugation to rapidly clear bacterial lysates post-alkaline lysis and 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 automated fluorescent DNA sequencing (typical reads exceed 800 bp), restriction enzyme digestion, ligation, PCR, in vitro transcription, transformation, and other applications.
- Speed – Purification of plasmid DNA < 40 minutes
- Safety – No phenol/chloroform extractions
- Versatile – Spin and vacuum formats available
- High quality – DNA is suitable for a variety of downstream applications
For Research Use Only. Not for use in diagnostic procedures.
|Downstream Application||Cloning, sequencing, transformation, PCR, restriction digestion, ligation, in vitro transcription etc.|
|Starting material||15-50 mL LB culture with OD600 between 2 and 3; or equivalent|
|Plasmid type||High-copy, low-copy, cosmid DNA|
|Processing mode||Manual (centrifugation or vacuum)|
|Throughput||1 - 24|
|DNA binding technology||Silica Midi Spin Column|
|Lysate clearance method||Filtration using Lysate clearance syringe|
|Processing time||<40 minutes|
|Yield||100-250 µg for high copy-number; 10-50 µg for low copy-number|
|Special note||Lysate clearance syringe for rapid sample processing|
Protocol and Resources
Product Documentation & Literature
Agarose Gel Electrophoretic analysis of purified plasmid
Figure 1. Plasmid DNA was isolated from eight 30 mL bacterial cultures grown 16 hours in LB medium using the E.Z.N.A.® FastFilter Plasmid Midi Kit and eluted in 300 µL elution buffer. Plasmid DNA (1% of total purified DNA) was analyzed on a 0.8% agarose gel. Lanes 1-8 represent plasmid DNA from 8 different bacterial cultures and Lane M represents the 1 kb DNA ladder.