<abstract xmlns="http://www.w3.org/1999/xhtml">

<p>DNA isolation is a fundamental technique for all molecular biology laboratories. Depending on the plant species, DNA isolation can be challenging. In particular, adapted protocols rarely exist for tree species which are not used as standard model organisms. Here, we describe a flexible DNA isolation protocol that works for 59 tree species in a modular system. It is based on an ATMAB-containing extraction buffer to which proteinase K and/or boric acid are added, depending on the plant species. Subsequent purification steps include one or two precipitations with dichloromethane and, depending on the tree species, an optional sodium acetate precipitation. Using leaf material of a hybrid poplar clone from <italic>in vitro</italic> culture, it was determined that higher amounts of DNA could be isolated with this material than from field leaves. Starting from leaf material, DNA isolation for difficult cases was achieved with cambium or root tissue. This protocol was used to extract DNA for subsequent PCR amplification. Markers for cpDNA, mtDNA, and genomic DNA were used for standardized testing.</p>
</abstract>

DNA isolation is a fundamental technique for all molecular biology laboratories. Depending on the plant species, DNA isolation can be challenging. In particular, adapted protocols rarely exist for tree species which are not used as standard model organisms. Here, we describe a flexible DNA isolation protocol that works for 59 tree species in a modular system. It is based on an ATMAB-containing extraction buffer to which proteinase K and/or boric acid are added, depending on the plant species. Subsequent purification steps include one or two precipitations with dichloromethane and, depending on the tree species, an optional sodium acetate precipitation. Using leaf material of a hybrid poplar clone from in vitro culture, it was determined that higher amounts of DNA could be isolated with this material than from field leaves. Starting from leaf material, DNA isolation for difficult cases was achieved with cambium or root tissue. This protocol was used to extract DNA for subsequent PCR amplification. Markers for cpDNA, mtDNA, and genomic DNA were used for standardized testing.

Flexible DNA isolation procedure for different tree species as a convenient lab routine

Silvae Genetica

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

DNA isolation is a fundamental technique for all molecular biology laboratories. Depending on the plant species, DNA isolation can be challenging. In...

Flexible DNA isolation procedure for different tree species as a convenient lab routine

Publicado en línea: 01 jul 2022

Páginas: 20 - 30

DOI: https://doi.org/10.2478/sg-2022-0003

© 2022 Tobias Bruegmann et al., published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.