The successful implementation of next-generation sequencing techniques in plant and woody tissues depends on the quality of initial starting material. This study demonstrated the use of a modified protocol that enabled the simultaneous extraction of both genomic DNA and total RNA from recalcitrant woody material. The geneticmaterial obtained by this protocol is of high quality and can be directly used in downstream analysis (e.g., next-generation sequencing). This protocol is particularly useful not only when the initial plant material is limited but also when genomic DNA features (e.g., methylation) have to be compared with the total RNA (e.g., gene expression). For such studies, the extraction from the same materials is highly preferred to minimize sample variation.
Lay abstract: The advancement of next-generation sequencing techniques has greatly facilitated almost all branches of biological and life-science studies. To guarantee reliable results and conclusions, high-quality starting genetic material (genomic DNA and total RNA) is essential. Our modified protocol, which is based on existing methods, enabled the efficient simultaneous extraction of genomic DNA and total RNA from the same recalcitrant woody sample. Thereby, it greatly minimizes the amount of initial plant materials needed for a specific study.
Fields of Science
- 4112 Forestry
- genomic DNA extraction
- next-generation sequencing studies
- total RNA extraction
- wood samples