An efficient method for total RNA extraction from peanut seeds |
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Authors: | C Huang J F Picimbon H Q Li Z Li Q Liu W Liu |
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Institution: | 1.Key Laboratory of Crop Genetic Improvement and Biotechnology, High-Tech Research Center,Shandong Academy of Agricultural Sciences,Jinan, Shandong,P.R. China;2.Key Laboratory of Crop Genetic Improvement and Biotechnology, Huanghuaihai,Ministry of Agriculture,Jinan,P.R. China;3.Functional Genomics and Proteomics of Chemical Ecology, High-Tech Research Center,Shandong Academy of Agricultural Sciences,Jinan, Shandong,P.R. China |
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Abstract: | Most of conventional RNA extraction methods failed to extract highly pure and integral RNA from peanut seeds because peanut
seeds are extremely rich in lipids, proteins, polysaccharides, and phenolic compounds. Here, we describe a new method, named
Peanut Improved Modified RNA extraction method (PIMRNAext), using SDS, Tris-saturated phenol, NaCl, and sarkosyl during the
extraction process, which are particularly successful for total RNA extraction from lipid- and polysaccharide-rich materials.
The proposed PIMRNAext method is simple and fast. It requires only conventional reagents and is completed within 2 h. Using
PIMRNAext gives very good yields of high quality peanut RNA. This method is about ten times more efficient than conventional
methods, and the RNA produced by it is compatible with further molecular biology experiments, such as RT-PCR. We propose to
use the PIMRNAext method to extract RNA from peanuts and peanut-like plant species not only for RT-PCR, but also for most
molecular biology techniques that need copies of pure RNA, such as microarray or cDNA library construction. |
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