African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12502

Full Length Research Paper

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

Zewei An1,2#, Qitong Wang1,2#, Yanshi Hu2, Yanhong Zhao3, Yacao Li1, Han Cheng2, Huasun Huang2*
  1College of Agriculture, Hainan University, Haikou, Hainan 570228, China . 2State Centre for Rubber Breeding and Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan 571737, China. 3College of Life Sciences, Ludong University, Yantai, Shangdong 264025, China.
Email: [email protected]

  •  Accepted: 10 April 2012
  •  Published: 15 May 2012

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 b-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 wordsHevea brasiliensis, polyethylene glycol (PEG), nucleic acid, co-extraction, higher plants.