4.3 Article

Effective strategies for isolating DNA from members of Asteraceae with high concentrations of secondary metabolites

期刊

BIOTECHNIQUES
卷 72, 期 3, 页码 85-89

出版社

FUTURE SCI LTD
DOI: 10.2144/btn-2021-0050

关键词

CTAB; DNA extraction; secondary metabolites; Silphium; sodium metabisulfite

资金

  1. Perennial Agriculture Project
  2. University of Colorado

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This article describes a method for extracting high-quality DNA from plant tissue containing secondary metabolites, and provides experimental evidence in the Asteraceae family. The results show that the modified CTAB method on young leaves yielded the highest DNA quantity and sufficient sample purity. There was no significant difference observed between fresh and lyophilized samples, providing a good foundation for DNA extraction from difficult plant samples.
METHOD SUMMARY A protocol for obtaining high-quality DNA extracted from plant tissue of species known to have interfering secondary metabolites is described. 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 - specifically, phenolic compounds - on the extraction of DNA from Silphium spp. This genus is 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 cetyl trimethylammonium bromide (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.

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