Overview
The E.Z.N.A.® Endo-Free Plasmid Maxi 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. 50-200 mL of bacterial cultures in LB medium typically produces 0.6-1.2 mg of high copy number plasmid DNA. Up to 500 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 <>
- Safe – No phenol/chloroform extractions
- Versatile – Spin and vacuum formats available
- High-quality – Transfection grade, low-endotoxin plasmid DNA (< 1="">
Specifications
For Research Use Only. Not for use in diagnostic procedures.
| Features | Specifications |
|---|---|
| Downstream Application | Transfection, gene therapy, cloning, in vitro Transcription, Next-GenerationSequencing, PCR, Transformation, restriction digestion, ligation |
| Starting material | 50-200 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 Maxi Spin Column |
| Lysate clearance method | Filtration using Lysate clearance syringe |
| Processing time | <45> |
| Yield | 600-1200 µg for high copy-number; 50-300 µg for low copy-number |
| Special note | Includes endotoxin removal step |
Protocol and Resources
Product Documentation & Literature
PROTOCOL
D6915 D6926 Endo-Free Plasmid Midi Maxi Kit
SDS
D6926 SDS
SALES SHEET
Product Data
Figure 1 Title
Figure 1. pEGFP-N1 plasmid was purified from 200 mL DH5α cultures harboring the plasmid using Omega Bio-tek’s E.Z.N.A.® Endo-Free Plasmid Maxi Kit following manufacturer’s recommended protocol. Plasmid DNA was eluted in 2 mL volume and quantified using Thermo Scientific’s NanoDrop™ 2000c system. The Endotoxin concentration was estimated based on a Limulus Amoebocyte Lysate (LAL) assay.
