Protocol for isolation of total RNA from mouse whole cochlea using organic solvents

TL;DR

This paper provides a detailed protocol for extracting RNA from mouse cochleae using organic solvents for gene expression studies.

Contribution

A new protocol for RNA isolation from mouse cochleae across a wide age range using organic solvents is introduced.

Findings

The protocol enables RNA isolation suitable for reverse transcription and PCR.

RNA quality is suitable for downstream applications across mice aged 2–25 weeks.

Abstract

Determining gene expression changes in cochlear components is important for understanding the mechanisms underlying hearing loss. Here, we present a protocol for the isolation of total RNA from mouse cochleae across a wide range of ages (2–25 weeks old) using organic solvent extraction. We describe the steps for obtaining the entire cochlea from mice and isolating high-quality total RNA suitable for further reverse transcription and PCR.

For complete details on the use and execution of this protocol, please refer to Rías et al.1

•Protocol for RNA isolation from mouse whole cochlea•Procedure for cochlea homogenization•Steps for aqueous phase obtention and RNA precipitation•Guidance for RNA quality testing and quantification

Protocol for RNA isolation from mouse whole cochlea

Procedure for cochlea homogenization

Steps for aqueous phase obtention and RNA precipitation

Guidance for RNA quality testing and quantification

Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Determining gene expression changes in cochlear components is important for understanding the mechanisms underlying hearing loss. Here, we present a protocol for the isolation of total RNA from mouse cochleae across a wide range of ages (2–25 weeks old) using organic solvent extraction. We describe the steps for obtaining the entire cochlea from mice and isolating high-quality total RNA suitable for further reverse transcription and PCR.

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