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RNA Isolation from Embryonic Zebrafish and cDNA Synthesis for Gene Expression Analysis
Published on: August 7, 2009
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Modified CTAB and TRIzol protocols improve RNA extraction from chemically complex Embryophyta.
Ingrid E Jordon-Thaden1, Andre S Chanderbali2, Matthew A Gitzendanner2
1Department of Biology, University of Florida, Gainesville, Florida 32611 USA ; University and Jepson Herbaria, University of California Berkeley, Berkeley, California 94720 USA.
Applications in Plant Sciences
|May 22, 2015
Summary
This study details effective RNA extraction protocols for diverse plant species, including woody and aromatic varieties. The TRIzol and TURBO DNA-free kit method is recommended for high-quality RNA suitable for sequencing.
Area of Science:
- Plant Molecular Biology
- Biotechnology
- Genomics
Background:
- RNA extraction is crucial for plant research, but chemically complex taxa pose challenges.
- Developing robust protocols is essential for accurate gene expression analysis.
Purpose of the Study:
- To present optimized RNA extraction protocols for diverse plant groups.
- To identify an effective method for chemically complex plant species.
Main Methods:
- Evaluated three RNA extraction methods on 91 plant taxa.
- Methods included TRIzol/TURBO DNA-free kits with sarkosyl, CTAB/TRIzol combinations, and CTAB/QIAGEN RNeasy kits.
Main Results:
- The TRIzol/sarkosyl/TURBO DNA-free kit method proved effective for 91 diverse plant taxa.
- Bench-ready protocols were developed for immediate application.
Conclusions:
- TRIzol supplemented with sarkosyl and the TURBO DNA-free kit is a robust method for RNA extraction from 100 mg of land plant leaf tissue.
- The developed protocols yield RNA of suitable quality, quantity, and stability for transcriptome sequencing.
