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从含有高浓度次生代谢物的菊科植物成员中分离DNA的有效策略。

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

作者信息

Bailey Dustin W, Attia Ziv, Reinert Stephan, S Hulke Brent, Kane Nolan C

机构信息

Ecology & Evolutionary Biology Department, University of Colorado, Boulder, CO 80309, USA.

Division of Biochemistry, Department of Biology, Friedrich-Alexander-University Erlangen-Nuremberg, 91058, Erlangen, Germany.

出版信息

Biotechniques. 2022 Mar;72(3):85-89. doi: 10.2144/btn-2021-0050. Epub 2022 Feb 7.

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 - specifically, phenolic compounds - on the extraction of DNA from spp. This genus is in the Heliantheae tribe of Asteraceae that also includes sunflower (). Three methods were attempted in , 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.

摘要

菊科是最大的植物科,但在基因组水平上却是研究最少的科之一。我们的研究探索了减少次生代谢物(特别是酚类化合物)对从 spp. 中提取DNA影响的实用方法。该属植物属于菊科向日葵族,向日葵()也在该族中。在实验中尝试了三种方法,所用叶片样本的条件和年龄各不相同。对幼叶采用改良的十六烷基三甲基溴化铵(CTAB)法可获得最佳的DNA产量,且样本纯度足够。对于任何提取方法或叶片年龄,新鲜样本和冻干样本之间均未观察到明显差异。这些结果为难提取的植物样本的DNA提取提供了良好的基础。

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