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Related Concept Videos

RNA-seq03:21

RNA-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...
Ribosome Profiling02:24

Ribosome 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.
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Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
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Related Experiment Video

Updated: May 26, 2026

Defining the Program of Maternal mRNA Translation during In vitro Maturation using a Single Oocyte Reporter Assay
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Defining the Program of Maternal mRNA Translation during In vitro Maturation using a Single Oocyte Reporter Assay

Published on: June 16, 2021

Gene expression in mouse oocytes by RNA-Seq.

Eric Antoniou1, Robert Taft

  • 1The Jackson Laboratory, Bar Harbor, ME, USA. Eric.antoniou@jax.org

Methods in Molecular Biology (Clifton, N.J.)
|December 7, 2011
PubMed
Summary
This summary is machine-generated.

This study details a protocol for preparing Illumina sequencing libraries from mouse oocytes and related cells. It enables genomic analysis even with limited starting material, overcoming challenges with small tissues or rare cell types.

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Area of Science:

  • Genomics
  • Molecular Biology
  • Next-Generation Sequencing

Background:

  • Second-generation sequencing technologies, like Illumina GAIIx, offer extensive genomic analysis capabilities.
  • Standard RNA sequencing protocols require substantial amounts of total RNA (≥1 μg).
  • Analyzing small tissues or rare cell types is often limited by low RNA yield.

Purpose of the Study:

  • To present a protocol for preparing Illumina sequencing libraries from limited biological samples.
  • To enable transcriptomic studies on mouse oocytes and associated cells.
  • To overcome RNA quantity limitations in next-generation sequencing.

Main Methods:

  • Protocol development for Illumina library preparation.
  • Utilizing minimal starting material from mouse oocytes.
  • Adaptation for mural cells, cumulus cells, and sorted/captured cells.

Main Results:

  • Successful generation of Illumina sequencing libraries from mouse oocytes.
  • Demonstration of protocol suitability for mural and cumulus cells.
  • Validation for flow-sorted or laser-captured cells.

Conclusions:

  • The developed protocol effectively prepares Illumina sequencing libraries from scarce biological samples.
  • This method expands the applicability of next-generation sequencing for transcriptomic analysis in challenging cell types.
  • Facilitates genomic studies on rare cells and small tissue samples.