Design, construction, and optimization of a monophase RNA extraction solution (Threezol) from plant samples

In recent years, the importance of nucleic acid molecules (DNA and RNA) as ideal biomarkers in genetic detection, identification and diagnosis is well-understood. Extraction and purification of DNA and RNA biomolecules are considered as the first step of genomics, functional genomics and transcriptomics experiments. To achieve a quick, efficient and simple RNA isolation kit, the development of monophase RNA extraction buffer (Phenol-based) namely Threezol (Riragene) was considered in this investigation. To evaluate Threezol solution, total RNA was extracted from different plant species, including wheat (Triticum aestivum L.), cucumber (Cucumis sativus L.), rice (Oryza sativa L.), Aeluropus (Aeluropus littoralis (Gouan) Parl.), tomato (Solanum lycopersicum L.), sesame (Sesamum indicum L.) and soybean (Glycine max (L.) Merr.). The quality and quantity of the extracted RNA were assessed using agarose gel electrophoresis, spectrophotometric method, and quantification analysis of ۱۸S rRNA by RT-qPCR method. Based on the ۲۸S/۱۸S rRNA ratio (۲۸S > ۱۸S) and OD۲۶۰/۲۸۰ ratio higher than ۱.۸, the extracted RNA from different plant samples had a similar quantity and quality with other commercial kits namely Trizol (Invitrogen) and RNAX Plus (Cinaclone). Based on RT-qPCR ۱۸S rRNA analysis in different plant samples, Ct less than ۱۸ was observed which could be an indication for the optimal quantity and quality of RNA extracted. The design, manufacture, and evaluation of these kits, while making the country self-sufficient in the production of this product, could be the starting point for the production and development of specific RNA/DNA extraction kits for animal and bacterial cells.

کلیدواژه ها:

Nucleic acid ، Extraction kit ، Threezol ، RT-qPCR ، Rragene