Location: Sunflower and Plant Biology ResearchTitle: Effective strategies for isolating DNA from members of Asteraceae with high concentrations of secondary metabolites
|BAILEY, DUSTIN - University Of Colorado|
|ATTIA, ZIV - University Of Colorado|
|REINERT, STEPHAN - University Of Colorado|
|KANE, NOLAN - University Of Colorado|
Submitted to: Biotechniques
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/13/2022
Publication Date: 2/7/2022
Citation: Bailey, D.W., Attia, Z., Reinert, S., Hulke, B.S., Kane, N.C. 2022. Effective strategies for isolating DNA from members of Asteraceae with high concentrations of secondary metabolites. Biotechniques. 72:85-89. https://doi.org/10.2144/btn-2021-0050.
Interpretive Summary: Extraction of DNA is a basic step in genetic analysis of organisms, but is often made difficult because of non-DNA compounds that contaminate the pure sample. This work attempted to find a practical method of DNA extraction for plants with large amounts of non-DNA contaminants. A method was discovered that consistently provided high quality DNA samples, and the resulting samples provided DNA sequence data of acceptable quality for further analysis. This will impact the study of many plants that previously had not been sequenced, including crop wild relatives like Silphium species.
Technical Abstract: The Asteraceae are the largest plant family, but among the least studied at the genome level. Our work investigated practical methods to reduce the influence of secondary metabolites on the extraction of DNA from Silphium spp., a genus in the Heliantheae tribe of Asteraceae that also includes sunflower (Helianthus annuus). Three methods were attempted in Silphium, with varying condition and age of the leaf sample. A modified CTAB method on young leaves resulted in the best DNA yield, with sufficient sample purity. No perceptible difference was observed between fresh and lyophilized samples for any extraction method or leaf age. These results provide an excellent basis for DNA extraction of difficult plant samples.