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Targeted DNA Methylation Analysis by Next-generation Sequencing
Published on: February 24, 2015
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MicroRNA Analysis Using Next-Generation Sequencing.
Monika Kolanowska1,2,3, Anna Kubiak1,2,3, Krystian Jażdżewski1,2,3
1Warsaw Genomics INC, Warsaw, Poland.
Methods in Molecular Biology (Clifton, N.J.)
|July 1, 2018
Summary
Next-generation sequencing (NGS) provides a comprehensive view of microRNA (miRNA) expression and sequence. This chapter details methods for RNA extraction and library preparation for miRNA analysis across various sample types.
Area of Science:
- Molecular Biology
- Genomics
- Biotechnology
Background:
- Next-generation sequencing (NGS) is a powerful tool for analyzing microRNA (miRNA) expression and sequence.
- Understanding the comprehensive miRNA landscape requires high-quality RNA extraction and library preparation.
Purpose of the Study:
- To describe methods for RNA extraction from diverse biological samples for miRNA analysis.
- To outline the principles of microRNA library preparation and sequencing compatible with various platforms.
Main Methods:
- RNA extraction from tissue specimens, serum, cytological slides, and FFPE samples.
- MicroRNA library preparation protocols.
- Sequencing using Illumina NextSeq 500 platform (principles adaptable to others).
Main Results:
- Detailed protocols for obtaining high-quality RNA suitable for NGS.
- Established methods for preparing microRNA libraries for sequencing.
- Demonstrated compatibility of principles across different sequencing technologies.
Conclusions:
- Effective RNA extraction and library preparation are crucial for comprehensive miRNA profiling using NGS.
- The described methods provide a robust framework for miRNA analysis from various sample types.
- The presented principles ensure adaptability to different sequencing platforms for broad applicability.
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