Plasmid MiniPrep Kit
{tab=Overview}
Plasmid DNA from bacterial cultures grown overnight was prepared with the Norgen Plasmid MiniPrep Kit and compared with two other commercial kits. The yield for each of the triplicates is shown on the graph (N1 to N3 for Norgen, and Y1 to Y3 and Z1 to Z3 for competitor kits).
Sequencing of purified plasmid DNA as a template resulted in an accuracy of 99% over a 950 bp contiguous sequence.
| Supplier |
|---|
| Norgen Biotek |
| Cat. no. |
| 1013300 |
| Kit size |
| 50 |
| Links |
| Norgen Biotek Protocol  Information Sheet MSDS Brochure |
Up to 25 ug plasmid DNA from 1.5 ml bacterial culture
Features
- High YieldRecovery is 5 to 25µg (depending on copy number)
- Versatile Plasmids up to 13kb can be purified
- High Column Capacity Bind up to 25µg of plasmid DNA.
- Fast Rapid spin-column format allows for the processing of multiple samples in 30 minutes.
- Complete Kits contain all columns and solutions required
The Norgen Plasmid Miniprep Kit is designed for the rapid purification of plasmid DNA from bacterial cultures. The kit allows for the removal of RNA, SDS, genomic DNA and other contaminants. The kit can also be used to prepare plasmid DNA in a concentrated form. The recovered DNA is of a high quality, and is suitable for a number of downstream applications including digestions, ligations, transformations and sequencing. The kit employs the use of a proprietary resin for DNA purification. This resin is an extremely inert and stable substance that is able to preferentially bind nucleic acids in the presence of high salts and low pH. Elution of the DNA is then dependent on a low salt concentration and higher pH conditions.
Purification is based on spin column chromatography using Norgens proprietary resin as the separation matrix. Norgens resin binds DNA under high salt concentrations and releases the bound DNA under low salt and slightly alkali conditions. The process for the isolation of plasmid DNA involves first pelleting 1.5 mL of an overnight culture of E. coli harbouring the plasmid of interest using centrifugation (please see the flow chart on page 4). The pellet is then resuspended in the provided Resuspension Buffer, which contains RNAase A. Lysis Solution is then added to the sample in order to assist in the lysis of the bacterial cells. Next, Binding Solution is added to the sample which will neutralize the sample and cause precipitation of the proteins and genomic DNA that is present. The resulting suspension is spun down, and the clarified lysate containing the plasmid DNA is then applied to a provided column through centrifugation. Norgens resin binds DNA in a manner that depends on ionic concentrations, thus the DNA will bind to the column while most of the RNA, proteins and other contaminants will either flowthrough or be retained on top of the resin. The bound DNA is then washed twice using the provided Wash Solution in order to remove any remaining impurities, and the purified PCR product is eluted with the Elution Buffer.
| Column Binding Capacity | 25 µg |
|---|---|
| Size of Plasmids Purified | Up to 13,000 bp |
| Average Yield from 1.5 mL of Culture | Up to 20 µg |
| Time to Complete 10 Purifications | 30 minutes |
High DNA Yield from Small Cultures (image principale)
Plasmid DNA from bacterial cultures grown overnight was prepared with the Norgen Plasmid MiniPrep Kit and compared with two other commercial kits. The yield for each of the triplicates is shown on the graph (N1 to N3 for Norgen, and Y1 to Y3 and Z1 to Z3 for competitor kits). High-Accuracy Sequencing of DNA Product
Sequencing of purified plasmid DNA as a template resulted in an accuracy of 99% over a 950 bp contiguous sequence.