Overview
The E.Z.N.A.® Plasmid Mini Kit I is designed to isolate up to 25 µg of high-quality plasmid DNA from 1-5 mL bacterial cultures in less than 30 minutes. Plasmid DNA purification follows the alkaline-lysis method and is simplified with HiBind® Mini Column technology into three quick steps: Bind, Wash, and Elute. Purified plasmid DNA is immediately ready for a wide variety of downstream applications such as routine screening, restriction enzyme digestion, transformation, PCR and DNA sequencing.The E.Z.N.A.® Plasmid Mini Kit is available in two formats – Q-spin (D6942) and V-spin (D6943). D6942 (Q-spin) features columns that are capless whereas D6943 (V-spin) includes columns that have a cap attached. The columns are otherwise identical in use and application and can be used in both vacuum or centrifugation protocols.
- Rapid – Purification of plasmid DNA in less than 30 minutes
- Safe – 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 | 1-5 mL LB culture |
| Plasmid type | High-copy, low-copy, cosmid DNA |
| Processing mode | Manual (centrifugation or vacuum) |
| Throughput | 1-24 |
| DNA binding technology | Silica Mini Spin Column |
| Lysate clearance method | Centrifugation |
| Processing time | <30> |
| Yield | 15-25 µg for high copy-number; 0.1-5 µg for low copy-number |
Kit Components
| Item | Available Separately |
|---|---|
| HiBind® DNA Mini Columns | View Product |
| 2 mL Collection Tubes | View Product |
| 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
D6942 D6943 D6945 Plasmid DNA Mini Kits I and II
SDS
D6942 SDS
SALES SHEET
Product Data
Plasmid DNA Yield and Purity from 4 mL Bacterial Culture using E.Z.N.A.® Plasmid Mini Kit I and a competing product from company Q
Figure 1. pGEM plasmid was purified from 4 mL DH5α cultures harboring the plasmid and eluted in 50 µL volume using kits from Omega Bio-tek and Company Q according to manufacturer’s recommended protocols. Plasmid DNA concentration was determined by optical density measurements using Thermo Scientific’s NanoDrop™ 2000c system.
Agarose Gel Electrophoretic analysis of purified plasmid
Figure 2. 5 µL of purified pGEM plasmid was analyzed on a 1% Agarose gel. Plasmid was isolated from 1 mL DH5α cultures harboring the plasmid using Omega Bio-tek’s E.Z.N.A.® Plasmid Mini Kit I.
DNA yield and quality using E.Z.N.A.® Plasmid Mini Kit I
Table 1. pGEM plasmid was purified from 1 mL DH5α cultures harboring the plasmid and eluted in 50 µL volume. Plasmid DNA concentration was determined by optical density measurements using Thermo Scientific’s NanoDrop™ 2000c system.
Endotoxin Level of Purified Plasmid DNA using E.Z.N.A.® Plasmid Mini Kit I
Table 2. Plasmid DNA purified with E.Z.N.A. Plasmid Mini Kit I was used in a 5 µL Sanger sequencing reaction. DNA was analyzed on an Applied Biosystems 3730XL.
Endotoxin Level of Purified Plasmid DNA using E.Z.N.A.® Plasmid Mini Kit I
Table 3. Endotoxins in plasmid DNA preps. Plasmid DNA was isolated from 0.8 mL LB cultures following each manufacturer’s recommended protocols. Endotoxin levels were determined with Thermo Scientific’s Pierce LAL Chromogenic Endotoxin Quantitation Kit.
Citations
- Guo, X., Xia, X., Tang, R., Zhou, J., Zhao, H., Wang, K. (2008). Development of real-time PCR method for Firmicutes and Bacteroidetes in faeces and its application to quantify intestinal population of obese and lean pigs. Letters in Applied Microbiology, 47: 367-373. http://dx.doi.org/10.1111/j.1472-765X.2008.02408.x
- Wang, H., Liu, X., Liu, S., Yu, Y., Lin, J., Pang, X., Zhao, J. (2011). Development of a markerless gene replacement system for Acidithibacillus ferroxidans and construction of a pfkB mutant. Applied nvironmental Microbiology, 78 (6): 1826-1835. doi: 10.1128/AEM.07230-11.
- Qi, H., Xiang, Z., Han, G., Yu, B., Huang, Z. Chen, D. (2011). Effects on different dietary protein sources on cecal microflora in rats. Africal Journal of Biotechnology, 10 (19): 3704-3807. doi: 10.5897/AJB.2677.
- Farnelid, H., Bentzon-Tilia, M., Andersson, A., Bertilsson, S., Jost, G., Labrenz, M., Jurgens, K., Riemann, L. (2013). Active nitrogen-fixing heterotrophic bacteria at and below the chemocline of the central Baltic Sea. The ISME Journal, 7: 1413-1423. doi: 10.1038/ismej.2013.26.
- Verhaegen, Y., Parmentier, K., Swevers, L., Renders, E., Rouge, P., de Coen, W., Cooreman, K., Smagge, G. (2011). The heterodimeric ecdysteroid receptor complex in the brown shrimp Crangon crangon: EcR and RXR isoform characteristics and sensitivity towards the marine pollutant tributyltin. General and Comparative Endocrinology,172: 158-169. doi: 10.1016/j.ygcen.2011.02.019.
- Li, R., Takala, T., Qiao, M., Xi, M., Saris, P. (2011). Nisin-selectable food-grade secretion vector for Lactococcis lactis. Biotechnology Letters, Springer Verlag, 33 (4): 797-803. doi: 10.1007/s10527-010-0503-6.
