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DNA Isolation01:24

DNA Isolation

DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...

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Back to Old Books Toward Affordable Research: Homemade Phenol-Based Reagent for Triphasic RNA Purification.

Mahmoud M Abdelfattah1, Yasmine Hosny2, Nadia A Elkady3

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Biochemical Genetics
|September 25, 2023
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Summary

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.

Keywords:
Acidic phenolEukaryotic cellsHomemade purificationPhenol-based reagentRNA

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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.