Simple and cheap method of RNA isolation from plant material

  • Д. Д. Буй
  • Я. В. Пирко
  • Я. Б. Блюм

Abstract

Aim. RNA isolation is the first essential step for molecular biological experiments. Number of different chemical methods of RNA extraction and purification are known. Even so, the high cost and complexity of many of these methods is an obstacle to their use in the regular laboratore practice. A new cheap modified method of RNA isolation is proposed. Methods. RNA isolation was conducted out using three different methods of extraction, including proposed by authors. Electrophoresis of obtained RNA samples was done. RNA, isolated from wheat Triticum aestivum, was used for real-time RT-PCR amplification of ubiquitin gene. Results. Proposed method of RNA extraction allows isolate RNA, which is good enough to be used in RT-qPCR. RNA elerctophoregrams in this case are similar to those, obtained with usage of commercial kits. The yield of RNA reaches 15–20 µg from 100 mg of plant tissue (leaf). Conclusions. The main advantages of proposed method are low cost and simple fast protocol. The main disadvantage is low stability of ready solution, resulting in frequent need to prepare a new one.

Keywords: RNA isolation, guanidinium thiocyanate.