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Updated: May 24, 2026
09:29
A Complete Pipeline for Isolating and Sequencing MicroRNAs, and Analyzing Them Using Open Source Tools
Published on: August 21, 2019
Deep-sequencing protocols influence the results obtained in small-RNA sequencing.
Joern Toedling1, Nicolas Servant, Constance Ciaudo
1Institut Curie, Paris, France.
Plos One
|March 3, 2012
Summary
Second-generation sequencing reveals microRNA and repetitive element small RNA expression. Library preparation protocols significantly impact results, necessitating consistent methods for accurate biological comparisons.
Area of Science:
- Molecular Biology
- Genomics
- Next-Generation Sequencing
Background:
- Second-generation sequencing is crucial for small RNA analysis.
- The influence of sequencing platforms and library preparation on small RNA data is understudied.
Purpose of the Study:
- To compare small RNA sequencing results using different platforms and protocols.
- To evaluate the impact of technical variations on microRNA and repetitive element small RNA expression.
Main Methods:
- Generated small RNA libraries from embryonic stem cells.
- Utilized three major second-generation sequencing technologies.
- Analyzed microRNA and repetitive element small RNA expression.
Main Results:
- Libraries showed good correlation across sequencing platforms.
- Qualitative and quantitative variations were observed based on the protocol.
- Protocol choice influenced the detection of microRNAs and repetitive element small RNAs.
Conclusions:
- Sequencing platform and library preparation protocols affect small RNA quantification.
- Consistent methodology is essential for reliable comparisons between biological samples.
- Standardization of protocols is recommended for reproducible small RNA research.
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