Emerging methods in botanical DNA/RNA extraction

Mitchell, Nora; McAssey, Edward V.; Hodel, Richard G. J.

doi:10.1002/aps3.11530

Cited by 1 publication

(1 citation statement)

References 29 publications

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“…Isolating high-quality RNA is essential for accurate downstream analyses, including gene expression studies, microarray analysis, RT-PCR, and RNA-Seq [15,26,27]. Obtaining a high-quality and quantity of RNA can be a significant challenge, and it often requires careful consideration of several factors [28]. The secondary metabolites, polysaccharides, and polyphenolic compounds present in conifers [29] can interfere with the RNA isolation process, leading to degraded RNA and resulting in low RNA yield and poor quality [5,8].…”

Section: Resultsmentioning

confidence: 99%

An Optimized and Cost-Effective RNA Extraction Method for Secondary Metabolite-Enriched Tissues of Norway Spruce (Picea abies)

Singh,

Naseer,

Sellamuthu

et al. 2024

Plants

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Since the development of next-generation sequencing techniques and with the growing interest in transcriptomic studies, there is a demand for high-throughput RNA extraction techniques. General RNA extraction protocols are unreliable when it comes to the quality and quantity of isolated RNA obtained from different tissue types of different plant species. Despite Norway spruce (Picea abies) being one of the most significant and commercially valuable tree species in European forests, only limited genetic research is available. In this study, we developed a cetyltrimethylammonium bromide (CTAB) protocol by modifying the original method. We compared this CTAB protocol with other widely used methods for extracting RNA from different tissues (needle, phloem, and root) of Norway spruce, known for its richness in polyphenols, polysaccharides, and secondary metabolites. The modified CTAB method proves to be superior to the kit-based and TRIzol-based methods for extracting RNA from the metabolite-rich tissues of Norway spruce, resulting in high RNA quality and integrity values (RIN~7–9). The modified CTAB RNA extraction method is rapid, cost-effective, and relatively simple in yielding the desired RNA quality from Norway spruce tissues. It is optimal for RNA sequencing and other downstream molecular applications.

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