Back to Old Books Toward Affordable Research: Homemade Phenol-Based Reagent for Triphasic RNA Purification
Mahmoud M Abdelfattah1, Yasmine Hosny2, Nadia A Elkady3
1Molecular Cancer Biology Group, Zoology Department, Faculty of Science, Ain Shams University, Cairo, Egypt.
Insights
Researchers developed a cost-effective, homemade RNA purification reagent using phenol and guanidine thiocyanate. This method provides comparable results to commercial kits, aiding research continuity during supply chain disruptions.
Area of Science:
- Biochemistry
- Molecular Biology
- Laboratory Science
Background:
- The COVID-19 pandemic disrupted global supply chains, impacting scientific research by delaying essential material and consumable deliveries.
- Developing countries face exacerbated challenges due to funding reallocation towards the COVID-19 crisis and increased reagent costs.
Purpose of the Study:
- To develop and validate a homemade, cost-effective reagent for RNA purification from eukaryotic cells and tissues.
- To address the reagent accessibility and cost issues faced by research laboratories, particularly in resource-limited settings.
Main Methods:
- Preparation of a homemade phenol-based RNA extraction reagent using saturated phenol (pH 4.3 adjusted with citrate buffer) and guanidine thiocyanate.
- Validation of the homemade reagent by purifying RNA from diverse biological samples including cell lines, tissues, and fungi.
- Comparison of RNA yield and quality from homemade versus commercial RNA extraction kits using identical starting materials.
Main Results:
- The homemade phenol-based reagent successfully purified RNA from various biological sources.
- RNA concentrations obtained using the homemade reagent were comparable to those from a commercial kit when processing the same number of cells.
- The cost-effectiveness of the homemade reagent was demonstrated, offering a viable alternative to commercial products.
Conclusions:
- The developed homemade phenol-based reagent is a cost-effective and efficient alternative for RNA purification.
- This reagent provides comparable performance to commercial kits, supporting uninterrupted research activities.
- The study highlights the feasibility of preparing essential laboratory reagents in-house to mitigate supply chain challenges and reduce research costs.
Abstract:
Covid-19 crisis did hit many socio-economic aspects in the whole world. In the scientific research, the problem is getting even worse, since most of materials and consumable are allocated to the health sector. Many research laboratories around the world have big delay in receiving their purchases to accomplish their research projects. In the developing countries, the situation is much more difficult, since most of the funding resources are directed to the Covid-19 crisis and there is a notable increase in reagents' prices. Therefore, the aim of the present study is to make a homemade reagents for RNA purification from eukaryotic cells/tissues. The homemade phenol-based RNA extraction reagents were prepared using saturated phenol pH 4.3 (adjusted by 0.5 M citrate buffer) and guanidine thiocyanate. To validate the phenol-based reagent, RNA was purified from different biological samples (cell line, tissues, and fungi) using homemade phenol-based versus a commercial one. Concentration of RNA samples extracted from the same number of cells were compared to assess the homemade preparation of phenol-based reagent. In conclusion, homemade phenol-based reagent is cost effective and comparable to the commercial one. Using homemade phenol-based, RNA extraction was successfully purified from different biological sources.
