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Published on: February 22, 2010
An effective method of RNA isolation from Fallopia multiflora tuberous roots.
Lei Chen1, Shu-Jing Sheng, Xue-Mei Tan
1Guangdong Provincial Key Lab of Biotechnology for Plant Development, College of Life Sciences, South China Normal University, Guangzhou, People's Republic of China.
Preparative Biochemistry & Biotechnology
|January 14, 2012
Summary
Isolating RNA from Fallopia multiflora tuberous roots is challenging due to high metabolite levels. A modified CTAB method successfully yields high-quality total RNA suitable for molecular applications.
Area of Science:
- Plant Molecular Biology
- Biochemistry
Background:
- Fallopia multiflora tuberous roots contain high levels of carbohydrates, phenolics, and secondary metabolites, complicating total RNA isolation.
- Existing RNA isolation protocols often yield poor results from such metabolite-rich plant tissues.
Purpose of the Study:
- To develop a modified protocol for high-quality total RNA isolation from Fallopia multiflora tuberous roots.
- To overcome challenges associated with secondary metabolites during RNA extraction.
Main Methods:
- A modified protocol was developed based on the traditional Cetyltrimethylammonium bromide (CTAB) method.
- The protocol was optimized for RNA extraction from Fallopia multiflora tuberous roots.
Main Results:
- The modified protocol successfully yielded high-quality and intact total RNA.
- RNA yield exceeded 0.15 mg/g fresh weight with an A260/A280 ratio of 1.9-2.0.
- The isolated RNA was suitable for downstream applications including RT-PCR, Northern hybridization, and cDNA library construction.
Conclusions:
- The developed protocol is effective for isolating high-quality total RNA from Fallopia multiflora tuberous roots.
- This method may be applicable to other plant species with similar metabolite profiles and tuberous roots.
