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An Efficient Method for the Isolation of Highly Purified RNA from Seeds for Use in Quantitative Transcriptome Analysis
Published on: January 11, 2017
An efficient method for purifying high quality RNA from wheat pistils
A Manickavelu1, Kumiko Kambara, Kohei Mishina
1Laboratory of Genetics and Plant Breeding, Chiba University, 648, Matsudo, Chiba 271-8510, Japan. agromanicks@rediffmail.com
Colloids and Surfaces. B, Biointerfaces
|December 5, 2006
Summary
This study presents a new method for extracting high-quality total RNA from wheat pistils, overcoming challenges posed by polysaccharides. The protocol significantly improves RNA yield and enables reliable downstream gene expression analysis.
Area of Science:
- Plant Molecular Biology
- Biochemistry
Background:
- Floral organs like wheat pistils contain high levels of polysaccharides that interfere with RNA extraction.
- Existing RNA extraction methods are often ineffective for these plant tissues.
Purpose of the Study:
- To develop a simple and effective protocol for total RNA extraction from wheat pistils.
- To improve RNA recovery and enable downstream molecular analyses.
Main Methods:
- Utilized Lithium chloride (LiCl) and phenol:chloroform:isoamylalcohol (PCI) for extraction.
- Employed microcentrifuge tubes with plastic pestles and liquid nitrogen for efficient grinding.
- Simplified procedures to minimize sample loss and maximize RNA recovery.
Main Results:
- Achieved a substantial increase in total RNA recovery (approximately 50%).
- Successfully performed reliable differential display by cDNA-AFLP using the extracted RNA.
- Demonstrated the method's efficacy for gene expression and regulation studies.
Conclusions:
- The developed method provides a robust approach for total RNA extraction from challenging plant floral tissues.
- This protocol is valuable for gene expression studies in wheat and potentially other plant species.
