
Nucleic Acid Purification
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® Plasmid DNA Miniprep Kit Protocol (NEB #T1010)
- Monarch® DNA Gel Extraction Kit Protocol (NEB #T1020)
- 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)
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- Troubleshooting Guide for DNA Purification
- 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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Legal Information
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For more information about commercial rights, please contact NEB's Global Business Development team at gbd@neb.com.
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.