Handling DNA
DNA cleanup
Cleanup of DNA is often a prerequisite for efficient downstream applications such as cloning, sequencing, microarray analysis, or amplification.The use of a dedicated kit for this application may be necessary, since kits can vary depending on the type of reaction and DNA fragment size (e.g., PCR products, gel extraction, enzymatic reactions, nucleotide removal, dye terminator removal) and the required elution volume.
Standard PCR cleanup only requires the removal of ~20 b oligos. In next-generation sequencing (NGS) library preparation, often, much larger primers — almost in the range of a PCR amplicon — must be removed, and PCR cleanup may not be sufficient. For this specialized “size selection” procedures, dedicated kits or PEG-based precipitation (8) are necessary.
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- Equipment and reagents required
- DNA precipitation
- Wash DNA pellet
- Resuspend DNA pellet
- Troubleshooting common problems in isopropanol DNA precipitation
- What are the advantages and disadvantages of isopropanol DNA precipitation?
- Role of sodium acetate in DNA precipitation
- Ethanol versus isopropanol DNA precipitation: What’s the difference?
- Post-precipitation DNA quantification and storage
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