Comparative study of DNA and RNA extraction methods for high-quality nucleic acid isolation from Cullenia exarillata

Isolation of high-quality DNA and RNA from plants with high polysaccharide and secondary metabolite content is typically difficult, particularly in the case of trees.  Metabolites commonly undergo co-precipitation with RNA and DNA, resulting in degradation of their quality. Cullenia exarillata leaf samples were subjected to various DNA and RNA extraction techniques, and the resulting data were compared and analysed. The isolation of high-quality DNA and RNA is crucial for the advancement of molecular and biotechnological techniques that aim to preserve the endemic status of C. exarillata and this research is to establish an efficient procedure for extracting DNA and RNA from C. exarillata. This method will make molecular and genomic research easier in forestry and conservation fields. In this research, we evaluated various methods for extracting high quality DNA and total RNA from C. exarillata tree, incorporating minor modifications in standard procedure. To acquire DNA and RNA of superior quality, a comparison was made between conventional DNA and RNA extraction methods and a variety of commercial kits that revealed the conventional technique yielded DNA samples of superior purity and concentration. It was discovered that combining modified commercial and conventional procedures yielded RNA with exceptionally high concentration and purity. The Agilent 2100 Bioanalyzer and NanoDrop spectrophotometer ensure the impeccable purity of the nucleic acids generated via these procedures. Additionally, the application of agarose gel electrophoresis unveiled unique bands. Further investigation was conducted to validate the purity and amplification of the DNA and RNA that were collected.  This study clarifies a method for extracting sufficient and high-quality amounts of DNA and RNA from C. exarillata; future research on this plant will greatly benefit from knowing this information.

Full article link: https://cellmolbiol.org/index.php/CMB/article/view/5841

Citation: Revathy S, Sabu K K, Anilkumar S (2025). Comparative study of DNA and RNA extraction methods for high-quality nucleic acid isolation from Cullenia exarillata A. Robyns. Cellular and molecular biology, 71(6), 54–61. https://doi.org/10.14715/cmb/2025.71.6.7

Small RNA manipulation in plants: Techniques and recent developments

The term “RNA silencing” refers to the processes that depend on small RNAs (approx. 18–24 nt) to regulate the mRNA expression in eukaryotes. In plants, small RNA regulation plays crucial roles in development, in responses to biotic or abiotic stress, in regulating metabolic pathways, in maintaining genome integrity, and in defense response mechanisms. In the last few years, small RNA manipulation has been used as a research tool to identify and validate gene expression, and studies now extensively focus on applying this technology in commercial applications, especially in medical therapeutics and flower color in ornamental plants. This chapter covers current progress on the application of RNA silencing to produce plants resistant to plant viruses, insects, and fungal pathogens, and also in applying these technologies toward the improvement of agricultural crops and medicinal plants. This chapter will also discuss the challenges, limitations, and biologic safety concerns related to small RNA manipulation in crops.

https://www.sciencedirect.com/science/article/pii/B9780128171127000183

High quality RNA extraction from small cardamom tissues rich in polysaccharides and polyphenols

Abstract

Due to the presence of a diverse array of metabolites, no standard method of RNA isolation is available for plants. We noted that polysaccharide and polyphenol contents of cardamom tissues critically hinder the RNA extraction procedure. Hence, we attempted several methods for obtaining intact mRNA and small RNA from various cardamom tissues. It was found that protocols involving a combination of commercial kits and conventional CTAB (cetyl trimethylammonium bromide) methods yielded RNA with good purity, higher yield, and good integrity. The total RNA isolated through this approach was found to be amenable for transcriptome and small RNA analysis through next-generation sequencing platforms.

Keywords

• RNA isolation; Next-generation sequencing (NGS); mRNA; miRNA; RIN value
• • http://dx.doi.org/10.1016/j.ab.2015.05.017 
  • Full paper: http://authors.elsevier.com/a/1RJrWlbACuNf
• 
  
• Analytical Biochemistry

• Fasiludeen Nadiya^a, 
• Narayanannair Anjali^a, 
• Appukuttannair Gangaprasad^b,
• Kalluvettankuzhy Krishnannair Sabu^a, , 

• ^a Biotechnology and Bioinformatics Division, Jawaharlal Nehru Tropical Botanic Garden & Research Institute (JNTBGRI), Thiruvananthapuram 695562, Kerala, India
• ^b Department of Botany, University of Kerala, Thiruvananthapuram 695037, Kerala, India

Received 22 May 2015, Accepted 28 May 2015, Available online 3 June 2015

Corresponding author

Kalluvettankuzhy Krishnannair Sabu - sabu@jntbgri.res.inBiotechnology and Bioinformatics Division, Jawaharlal Nehru Tropical Botanic Garden & Research Institute (JNTBGRI), Thiruvananthapuram 695562, Kerala, India.