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Isolation of Transcriptomic-Quality Total RNA from Mouse Spinal Cords
John V Stokes1,2, Ashleigh J Nicaise1,2,3, Christa M Frodella1,2
1Department of Comparative Biomedical Sciences, Mississippi State University, Mississippi State, Mississippi.
Current Protocols
|January 14, 2022
Summary
This protocol details spinal cord RNA isolation from mice, crucial for studying neuroinflammation and neurodegenerative diseases like multiple sclerosis. Optimized methods ensure high-quality RNA yield for transcriptomic analysis.
Area of Science:
- Neuroscience
- Immunology
- Molecular Biology
Background:
- Spinal cord analysis is key to understanding neuroinflammation and neurodegenerative diseases.
- Immune cell infiltration in the spinal cord is a hallmark of conditions like multiple sclerosis.
- Total RNA assessment provides insights into neuronal and inflammatory profiles and drug effects.
Purpose of the Study:
- To provide a detailed protocol for spinal cord explantation and homogenization.
- To demonstrate a method for optimal RNA isolation from mouse spinal cords.
- To enable transcriptomic analysis for neuroinflammation and neurodegeneration research.
Main Methods:
- Spinal cord isolation from the mouse spinal column using a specialized needle.
- Homogenization of the spinal cord tissue using liquid nitrogen.
- Assessment of RNA purity, quantification, and integrity.
Main Results:
- The protocol ensures efficient RNA isolation from mouse spinal cords.
- The method is applicable to both healthy and experimentally induced neuroinflammatory conditions (e.g., EAE).
- Successful implementation yields sufficient transcriptomic-quality RNA.
Conclusions:
- This protocol is essential for researchers studying spinal cord pathology.
- High-quality RNA isolation is critical for accurate analysis of neuroinflammation and neurodegeneration.
- The described technique supports advancements in understanding and treating neurological diseases.
