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African Journal of Biotechnology

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Co-extraction of high-quality RNA and DNA from rubber tree (Hevea brasiliensis)

Zewei An, Qitong Wang, Yanshi Hu, Yanhong Zhao, Yacao Li, Han Cheng, Huasun Huang

Abstract


High-quality nucleic acids are the basic requirement for performing genomic research. A reliable and efficient method was developed for co-extracting high-quality DNA and RNA from rubber tree (Hevea brasiliensis) in this study. Polyethylene glycol (PEG) and cetyltrimethylammonium bromide (CTAB) extraction buffer with high concentrations of polyvinylpyrrolidone (PVP) and β-mercaptoethanol was used in this study. The results show that 3.2% polyethylene glycol 8000 is the optimal concentration for successful separation of DNA and RNA. Spectrophotometric determination (A260/A280 and A260/A230 ratios), agarose electrophoresis analysis and reverse transcription (RT-PCR) of isolated nucleic acids indicate that high-quality DNA and RNA were extracted by this method. The general applicability of this method was also evaluated, and the results show that it was suitable for a variety of plants.

Key words: Hevea brasiliensis, polyethylene glycol (PEG), nucleic acid, co-extraction, higher plants.




http://dx.doi.org/10.5897/AJB12.136
AJOL African Journals Online