The purification of plasmid DNA from bacterial cells is an important step in the cloning workflow. During plasmid purification, bacterial cells are lysed, freeing DNA and other cellular components from the cell wall. Cellular components are then removed, and the DNA-containing lysate is processed to further remove contaminants separate the plasmid DNA from the genomic DNA.
The Monarch Plasmid Miniprep Kit is a rapid and reliable method for the purification of high quality plasmid DNA from bacterial cultures. This method employs standard cell resuspension, alkaline lysis, and neutralization steps, with the additional benefit of color indicators at certain steps to easily monitor completion. Unique wash buffers ensure that salts, proteins, RNA and other cellular components are removed, allowing for the elution of concentrated, highly pure DNA. Elution in as little as 30 µl provides concentrated DNA for use in downstream applications, such as restriction enzyme digestion, DNA sequencing, PCR, other enzymatic manipulations and transfection of robust cell lines. To learn more, view performance data for the Monarch Plasmid Miniprep Kit.
Monarch Plasmid Miniprep Kits provide fast and reliable purification, reduced hands-on time and elution in low volumes.
FAQs for Plasmid Purification
- What is the maximum binding capacity of the Monarch Plasmid Miniprep Column?
- What is the smallest volume of elution buffer that can be used with the Monarch Plasmid Miniprep Kit?
- Which culture media do you recommend?
- Which strain(s) of E.coli do you recommend?
- What should I do differently when using a low copy plasmid?
- What factors affect recovery of plasmid DNA?
- Can I use DNA isolated form the Monarch Plasmid Miniprep Kit for transfection?
- What factors affect my (A260/A230) after doing a plasmid miniprep?
- What factors affect my (A260/A280) after doing a plasmid miniprep?
- What culture conditions do you recommend?
- I have used 5 ml of cell culture for plasmid isolation with the Monarch Plasmid Miniprep kit and I am obtaining low amounts of plasmid and/or contaminating gDNA. Why is this, and what are your suggestions to improve yield and purity?
- Are plasmids recovered using the Monarch Plasmid Miniprep Kit endotoxin free?
- Are Monarch spin columns compatible with Vacuum Manifolds?
- Can I use water to elute the DNA when using the Monarch Kits?
Protocols for Plasmid Purification
- Using Exonuclease V (RecBCD) to eliminate residual genomic DNA when purifying low copy plasmids with the Monarch® Plasmid Miniprep Kit
Monarch Nucleic Acid Purification Brochure
It’s time to transform your DNA and RNA purification experience! Learn about the advantages of choosing Monarch Nucleic Acid Purification Kits from NEB.
- Troubleshooting Guide for DNA Purification
- Dos and Don'ts of Plasmid Minipreps
- Usage Guidelines for the Monarch Plasmid Miniprep Kit (#T1010) When Working with Low Copy Plasmids
Other Tools & Resources
Advantages of the Monarch Plasmid Miniprep Kit
Advantages of the Monarch Plasmid Miniprep Kit
- Elute in as little as 30 μl
- Prevent buffer retention and salt carry-over with optimized column design
- Reduce hands on time with faster protocols and less spin time
- Monitor completion of certain steps using colored buffer system
- No need to add RNase before starting
- Easily label columns using tab and frosted surfaces
- Buffers and columns available separately
- Significantly less plastic used when compared with other kits
- Responsibly-sourced and recyclable packaging
- No hazardous materials fees
Specifications of the Monarch Plasmid Miniprep Kit
- Culture Volume: 1-5 ml, not to exceed 15 OD units.
- Binding Capacity: up to 20 μg
- Plasmid Size: up to 25 kb
- Typical Recovery: up to 20 μg. Yield depends on plasmid copy number, host strain, culture volume, and growth conditions.
- Elution Volume: ≥ 30 μl
- Purity: A260/280 and A260/230 ≥ 1.8
- Protocol Time: 10.5 minutes of spin and incubation time
- Compatible Downstream Applications: restriction digestion and other enzymatic manipulations, transformation, transfection of robust cells, DNA sequencing, PCR, labeling, cell-free protein synthesis, etc.
While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.
For more information about commercial rights, please contact NEB's Global Business Development team at firstname.lastname@example.org.
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
Learn how to do a plasmid miniprep using the Monarch Plasmid Miniprep Kit.
Optimize your plasmid miniprep results with these quick tips for using the Monarch Plasmid Miniprep Kit.
Learn how you can easily recycle all of the components in your Monarch Kits.