Isolation of RNA without toxic reagents
MSU scientists have developed a new method for isolating RNA from viruses, bacteria, plants and animals
MSU Press Service
The main task in isolating RNA has always been to get rid of its protein. This process is called deproteinization. To isolate RNA from viruses, bacteria and higher organisms, substances are used that cause the destruction of complex cellular and viral complexes in which RNA is located. For this purpose, the most frequent was the use of surfactants and phenol. Their combination makes it possible to isolate RNA from almost all viruses and cells of living organisms. At the same time, nucleic acids pass into an aqueous solution, and with each extraction, 80% of the protein is extracted into phenol.
In the late 80s. Pyotr Khomchinsky and Nicolet Sakhi proposed an effective single–stage treatment of cells with a solution of a mixture of guanidine thiocyanate, which causes the breakdown of RNA complexes with proteins, and phenol with chloroform (the brand name of the mixture is TRIzol) to isolate total RNA (cellular and viral). The reagents used in the method – guanidine thiocyanate, phenol and chloroform – are very toxic.
Due to the rapid development of biotechnologies, RNA isolation is becoming widespread. The environmental friendliness of RNA isolation methods comes to the fore.
Yuri Drygin, Doctor of Chemical Sciences, Head of the Laboratory of Nuclein-Protein Interactions at the A.N. Belozersky Moscow State University, proposed using ammonium trichloroacetate (THAA) for RNA isolation.
RNA preparations obtained from viruses, E.coli, plants and animal cells using THAA do not differ in physico-chemical properties from preparations isolated by previously known methods. Ammonium trichloroacetate (THAA) is not included in the International List of Toxic Substances, it is harmless to plants, does not affect animal and human cells in vitro and in vivo. This circumstance is especially important for the mass isolation of RNA, which is necessary, for example, for the diagnosis of viral infections in emergency circumstances: in case of mass infection of plants, animals and people (epiphytotics, epizootics and pandemics).
Virus RNA polymerase. Source: InfoCan
In a joint publication and an application for a grant from the RFBR, scientists from the A.N. Belozersky Research Institute of Physico-Chemical Biology of Moscow State University with colleagues from the Institute of Bioresource Technologies in the Himalayas (CSIR – Institute of Himalayan Bioresource Technology) are developing methods of isolation and detection of RNA based on the developed method in order to detect viruses and viroids of perennial fruit plants.
Article by Kondakova et al. RNA isolation with low toxic ammonium trichloroacetate was published in the European Journal of Biotechnology and Science.
Portal "Eternal youth" http://vechnayamolodost.ru
11.05.2016