The Journal of Horticultural Science & Biotechnology
Vol. 87 No: 4
A modified and inexpensive protocol for the rapid isolation of RNA from diverse plant species
DANA JAWDAT and INTISSAR KARAJOLI
The availability of an inexpensive and rapid procedure for total RNA isolation, specifically from recalcitrant plant species such as cotton, would enhance the efficiency of transcriptomic research.
This work describes the successful modification of a total RNA isolation procedure.
The modified procedure takes only 150 min to complete, with high yields and a cost reduction of 90% over available commercial kits on the market.
The procedure presented in this work follows the hot borate method with modifications in the reagents, tissue weight, incubation periods, and supernatant filtration steps used.
The modified procedure was used successfully to isolate total RNA from recalcitrant plant tissues high in phenolic compounds such as those from cotton, dicots (tobacco and potato), monocots (wheat and barley), and woody perennials (poplar), suggesting its possible wide use in many multi-disciplinary research laboratories.
The results showed that the isolated RNA had no inhibitory effects during subsequent applications such as RT-PCR.
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