Overview
The E.Z.N.A.® Plasmid Midi Kit utilizes the convenient spin-column format to deliver high-quality plasmid DNA following a simple “bind-wash-elute” procedure without need for expensive accessories. 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 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
Specifications
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 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 | Centrifugation |
| Processing time | <60> |
| Yield | 100-250 µg for high copy-number; 10-50 µg for low copy-number |
Kit Components
| 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 |
Protocol and Resources
Product Documentation & Literature
PROTOCOL
D6904 Plasmid DNA Midi Kit
SDS
D6904 SDS
SALES SHEET
Product Data
Yield and quality of plasmid DNA purified using E.Z.N.A.® Plasmid Midi and Maxi Kits
Table 1. pGEM plasmid was purified from 50 mL DH5α cultures harboring the plasmid using Omega Bio-tek’s E.Z.N.A.® Plasmid Midi Kit following manufacturer’s recommended protocol. Plasmid DNA was quantified using Thermo Scientific’s NanoDrop™ 2000c system.
Citations
- Huang, Ally, et al. “BioBitsTM Explorer: A Modular Synthetic Biology Education Kit.” Science Advances, vol. 4, no. 8, 1 Aug. 2018, p. eaat5105, advances.sciencemag.org/content/4/8/eaat5105?intcmp, 10.1126/sciadv.aat5105. Accessed 1 June 2020.
- Liu, Ning, et al. “Cholecalciterol Cholesterol Emulsion Attenuates Experimental Autoimmune Myocarditis in Mice via Inhibition of the Pyroptosis Signaling Pathway.” Biochemical and Biophysical Research Communications, vol. 493, no. 1, 4 Nov. 2017, pp. 422–428, www.sciencedirect.com/science/article/pii/S0006291X17317643?casa_token=n1HisU6nWhYAAAAA:EtQ9hZAu9dkx7DKTaREOW6UpOhyBrxI94aV_OV5mY7Ul6apJDuN7QcmfQ46BVCIWHUMjyNCYslM, 10.1016/j.bbrc.2017.09.006. Accessed 1 June 2020.
- Seneviratne, Ayesh K., et al. “The Mitochondrial Transacylase, Tafazzin, Regulates AML Stemness by Modulating Intracellular Levels of Phospholipids.” Cell Stem Cell, vol. 24, no. 4, Apr. 2019, pp. 621-636.e16, 10.1016/j.stem.2019.02.020. Accessed 9 May 2020.
- Singh, Rashim Pal, et al. “Disrupting Mitochondrial Copper Distribution Inhibits Leukemic Stem Cell Self-Renewal.” Cell Stem Cell, 15 May 2020, www.sciencedirect.com/science/article/pii/S1934590920301491?casa_token=wHV9XQbteGIAAAAA:k6om0ZTifLELJJzXSk2XjBwrrWWCLBUbe3D6PnUTf1ax-283bQYtvGR1Z5xMQisbHvKQujvM0HA, 10.1016/j.stem.2020.04.010. Accessed 1 June 2020.
- Tippens, Nathaniel D., et al. “Transcription imparts architecture, function, and logic to enhancer units.” bioRxiv (2019): 818849.
