When isolating nucleic acids from cells, the first step is to disrupt the cell membrane to release the DNA. This is often done using chaotropic salts, alkaline denaturation, or detergents. Next, the soluble components of the cell lysate must be separated from the cellular debris and other insoluble materials. Centrifugation and vacuum filtration are frequently used to accomplish this separation, but magnetic clearing has gained wide acceptance more recently. Lastly, the nucleic acids must be separated from the other soluble materials in the lysate, including proteins.
Once the DNA is in solution, regardless of whether it originated from cells or from an enzymatic reaction, there are many different methods that can be employed to isolate it from other soluble components.
FAQs for Nucleic Acid Purification
- 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?
- Can I use the Monarch DNA & PCR Cleanup Kit to purify oligonucleotides and other short DNA fragments?
Protocols for Nucleic Acid Purification
- Monarch® DNA Gel Extraction Kit Protocol (NEB #T1020)
- Monarch® Plasmid DNA Miniprep Kit Protocol (NEB #T1010)
- Oligonucleotide Cleanup Using Monarch® PCR & DNA Cleanup Kit (5 μg) Protocol (NEB #T1030)
- Protocol for DNA Cleanup and Concentration Using the Monarch® PCR & DNA Cleanup Kit (5 μg) (NEB #T1030)
- Quick Protocol for DNA Cleanup and Concentration Using the Monarch® PCR & DNA Cleanup Kit (5 μg) (NEB #T1030)
- Quick Protocol for Monarch® DNA Gel Extraction Kit (NEB #T1020)
- Quick Protocol for Monarch® Plasmid Miniprep Kit (NEB #T1010)
- Quick Protocol for Oligonucleotide Cleanup Using the Monarch® PCR & DNA Cleanup Kit (5 μg) (NEB #T1030)
Five Steps to a Greener Lab: A roadmap to environmental action
The laboratory is a place of inspiration, curiosity, ingenuity, and often altruism. Researchers are, by and large, a thoughtful and deeply invested group, but the environmental and ecological consequences of life science research are not typically at the forefront of researchers’ minds when planning their investigations.
- Troubleshooting Guide for DNA Cleanup and Plasmid Purification
- Back to basics: Important things to keep in mind when purifying plasmids and DNA fragments
- Dos and Don'ts of Plasmid Minipreps
- Six Tips for a Perfect Gel Extraction
- Usage Guidelines for the Monarch Plasmid Miniprep Kit (#T1010) When Working with Low Copy Plasmids
Other Tools & Resources
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Learn how to do a plasmid miniprep using the Monarch Plasmid Miniprep Kit.
Learn how to extract DNA from agarose gels using the Monarch DNA Gel Extraction Kit.
Learn how to isolate DNA from your enzymatic reactions, including PCR, using the Monarch PCR & DNA Cleanup Kit (5 µg).
Optimize your plasmid miniprep results with these quick tips for using the Monarch Plasmid Miniprep Kit.
Optimize your DNA gel extractions with our quick tips for using the Monarch DNA Gel Extraction Kit.
Optimize your DNA isolation from PCR and other enzymatic reactions with our quick tips for using the Monarch PCR & DNA Cleanup Kit.
Learn how you can easily recycle all of the components in your Monarch Kits.