Related Experiment Video
Updated: Jun 21, 2026
15:21
RNA Isolation from Embryonic Zebrafish and cDNA Synthesis for Gene Expression Analysis
Published on: August 7, 2009
38.2K
Validation of RNA Extraction Methods and Suitable Reference Genes for Gene Expression Studies in Developing Fetal
Claudia Steinacher1, Dietmar Rieder2, Jasmin E Turner3
1Department of Otorhinolaryngology, Medical University Innsbruck, 6020 Innsbruck, Austria.
International Journal of Molecular Sciences
|March 13, 2024
Summary
High-quality RNA extraction from fresh fetal human inner ear tissue is crucial for gene expression studies. Fresh tissue in RNAlater yielded superior RNA, and four reference genes (B2M, HPRT1, GAPDH, GUSB) were validated for accuracy.
Area of Science:
- Developmental Biology
- Molecular Biology
- Genetics
Background:
- Accurate gene expression analysis relies on high-quality RNA.
- Proper reference gene selection is essential for reliable gene expression studies, especially in developmental contexts.
Purpose of the Study:
- To compare RNA extraction methods for fetal human inner ear tissue.
- To identify stable reference genes for gene expression studies in the developing human inner ear.
Main Methods:
- RNA was extracted from fresh tissue (RNAlater) and formalin-fixed paraffin-embedded (FFPE) tissue.
- Twelve reference genes were evaluated for expression stability in fetal inner ear samples (gestational weeks 11-19).
- Expression of target genes (OTOF, TECTA) was analyzed using validated reference genes.
Main Results:
- Fresh tissue stored in RNAlater yielded significantly higher RNA yield and quality compared to FFPE tissue.
- Four reference genes (B2M, HPRT1, GAPDH, GUSB) demonstrated stable expression across developmental stages.
- The selected reference genes did not affect the expression profile of OTOF and TECTA.
Conclusions:
- The choice of RNA extraction method critically impacts RNA quality and yield for gene expression studies.
- Validated reference genes are essential for accurate gene expression analysis in the fetal human inner ear development.
Related Concept Videos
03:21RNA-seq
RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...
02:15Experimental RNAi
RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...
02:24Ribosome Profiling
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...