E.Z.N.A.® Plasmid DNA Midi Kit
Price range: $0.00 through $715.50
- Speed – Purification of plasmid DNA in < 60 minutes
- Safety – No phenol/chloroform extractions
- Versatile – Spin and vacuum formats available
- High quality – DNA is suitable for a variety of downstream applications
The E.Z.N.A. Plasmid Midi and Maxi Kits can isolate up to 250 μg and 1.2 mg of plasmid DNA from 50 mL and 200 mL bacterial cultures, respectively. These kits use a modified alkaline lysis method to lyse the cells and separate genomic DNA from plasmid DNA. Cellular debris are removed by centrifugation, and the protocol follows a simple bind, wash, and elute procedure to deliver high-quality
plasmid DNA. The system not only eliminates the time-consuming isopropanol precipitation step required by gravity flow columns but also the need for expensive accessories. 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.
For Research Use Only. Not for use in diagnostic procedures.
| FEATURES | SPECIFICATIONS |
|---|---|
| Downstream Application | Cloning, sequencing, transformation, PCR, restriction digestion, ligation, in vitro transcription etc. |
| Starting material | 15-50 mL LB culture; Optimal Cell Mass (OD600 x mL culture) is 80-100. |
| 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 | Centrifugation |
| Processing time | <60 minutes |
| Yield | 100-250 µg for high copy-number; 10-50 µg for low copy-number |
| ITEM | AVAILABLE SEPARATELY |
|---|---|
| HiBind® DNA Mini Columns | View Product |
| 15 mL Collection Tubes | — |
| Solution I | View Product |
| Solution II | View Product |
| Solution III | View Product |
| HBC Buffer | View Product |
| DNA Wash Buffer | View Product |
| RNase A | View Product |
| Elution Buffer | View Product |
High-Quality Plasmid Purification
Figure 1. pGEM and pGL4.51 plasmids were purified from 50 mL DH5α cultures and eluted in 500 µL volume using Omega Bio-tek’s E.Z.N.A.® Plasmid DNA Midi Kit. Plasmid DNA concentrations were determined by optical density measurements using Thermo Scientific’s NanoDropTM 2000c system. All samples have ideal A260/A280 (~1.8) and A260/A230 (~2.0) absorbance ratios, indicating high-quality purified DNA.
Unfragmented Plasmid Purification
Figure 2. 200 ng purified pGEM and pGL4.51 plasmid were analyzed on a 0.8% Agarose gel. Plasmids were isolated from 50 mL DH5α cultures using Omega Bio-tek’s E.Z.N.A.® Plasmid DNA Midi Kit. Plasmids were digested with HF-ScaI to linearize.
Sequencing-Quality Plasmid Purification
| Plasmid Type | Sample | Trace Score | Continuous Read Length | QV20+ |
|---|---|---|---|---|
| pGEM | 1 | 56 | 982 | 983 |
| 2 | 56 | 958 | 963 | |
| 3 | 56 | 957 | 964 | |
| pGL4.51 | 1 | 57 | 954 | 955 |
| 2 | 55 | 948 | 951 | |
| 3 | 54 | 956 | 959 |
Table 1. pGEM and pGL4.51 plasmids were purified from 50 mL DH5α cultures harboring the plasmids and eluted in 500 µL volume using Omega Bio-tek’s E.Z.N.A.® Plasmid DNA Midi Kit. The plasmids were then used for Sanger sequencing. All samples display high continuous read length and QV20+ scores, indicating plasmids of high-quality isolated from this kit.