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Updated: Jun 4, 2026
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Discrimintion and Mapping of the Primary and Processed Transcripts in Maize Mitochondrion Using a Circular RT-PCR-based Strategy
Published on: July 29, 2019
T7-based RNA amplification for genotyping from maize shoot apical meristem
Kazuhiro Ohtsu1, Patrick S Schnable
1Department of Agronomy, Iowa State University, Ames, IA 50011, USA.
CSH Protocols
|March 2, 2011
Summary
This study presents a protocol for amplifying RNA from laser-captured tissues. This method enables SNP analysis and detection using expressed sequence tags derived from amplified RNA.
Area of Science:
- Molecular Biology
- Genomics
- Transcriptomics
Background:
- RNA offers a smaller, less repetitive target for genotyping compared to genomes.
- Laser capture microdissection (LCM) isolates specific tissue sequences, including rare transcripts.
- Standard LCM yields insufficient RNA for global expression analysis.
Purpose of the Study:
- To describe a protocol for amplifying RNA extracted from laser-dissected tissues.
- To enable SNP analysis and detection from limited RNA samples.
Main Methods:
- RNA extraction from laser-captured tissues and cells.
- Linear RNA amplification to increase yield.
- Conversion to complementary DNA (cDNA).
- 454 sequencing of cDNA to generate expressed sequence tags (ESTs).
Main Results:
- Successful amplification of RNA from microdissected samples.
- Generation of ESTs suitable for SNP analysis.
- Demonstration of RNA's utility in genotyping.
Conclusions:
- Amplified RNA from LCM is a viable source for SNP detection.
- This protocol enhances the application of RNA in genetic analysis.
- Facilitates genotyping of specific tissues and rare transcripts.
