Abstract
Research on bone diseases often requires investigation of bone gene expression. Isolating high-quality RNA is essential to obtain reliable and accurate gene expression data. In an effort to analyze the expression of genes related to osteoporosis in rat bones, we developed an improved method for extraction of high-quality RNA without the need for liquid nitrogen or specialized equipment. This method involved transitioning frozen bone tissues to a more pliable state with RNAlater ice and pulverization of the samples with a simple homogenizer, followed by a phenol-chloroform-based RNA extraction. Spectrophotometric analysis indicated high purity of the isolated RNA. Electrophoresis on agarose gel revealed two well-defined ribosomal RNA bands. Herein, we present a method that consistently yields RNA of high purity and integrity from rat bone.
Highlights
Research on bone diseases, including osteoporosis, metabolic bone disorders, and inherited syndromes, often requires investigation of bone gene expression
The conventional approach involves grinding frozen bone samples wrapped in foil using a mortar and pestle, pre-chilled with liquid nitrogen [1,2,3,4]
We present an improved method for RNA isolation from rat bone tissue without the need for liquid nitrogen or specialized equipment
Summary
Research on bone diseases, including osteoporosis, metabolic bone disorders, and inherited syndromes, often requires investigation of bone gene expression. We developed an improved method that consistently yields high-quality RNA from bone while eliminating the need for liquid nitrogen and specialized equipment. How to cite this article Poutoglidou F, Saitis A, Pourzitaki C, et al (March 10, 2021) An Improved Method for Isolating High-Quality RNA From Rat Bone at Room Temperature Without the Need for Specialized Equipment. Bone samples were transferred from -80°C storage to 2 mL Eppendorf tubes containing 1.5 mL of pre-chilled RNAlater ice (InvitrogenTM, Carlsbad, CA, USA) and stored at -20°C for at least 16 hours before RNA isolation. Once the initial overnight soak at -20°C was complete, bone samples were removed from RNAlater ice, sectioned into small pieces, weighted, and the desired amount (100 mg) was placed into homogenization tubes containing 1.5 mL of pre-chilled TRIzol Reagent (InvitrogenTM). The absence of “smearing” indicates high RNA quality (Figure 1)
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