Environmentally friendly method of RNA isolation

The diversity of organisms, tissues and cells is so great that, to date, no universal method for RNA extraction from these biological materials exist. The RNA isolation technique with a mix of guanidine thiocyanate, phenol, and chloroform is most widely used. Extraction and purification of RNA metho...

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Veröffentlicht in:Analytical biochemistry 2021-05, Vol.620, p.114113, Article 114113
Hauptverfasser: Drygin, Yuri F., Butenko, Konstantin O., Gasanova, Tatiana V.
Format: Artikel
Sprache:eng
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Zusammenfassung:The diversity of organisms, tissues and cells is so great that, to date, no universal method for RNA extraction from these biological materials exist. The RNA isolation technique with a mix of guanidine thiocyanate, phenol, and chloroform is most widely used. Extraction and purification of RNA methods using selling guanidinium-phenol (TRIzol)-based and silica-based column kits have limitations on toxicity, or RNA isolation, particularly for plants, and scaling. The agents' toxicity is particularly relevant when employing for mass analysis in practice while gaining RNA preparations during the pandemics, epizootics, and epiphytotic. In modern diagnostics of infections at the molecular level, powerful RT-PCR technology is used, which amplifies the detection of RNA pathogens by hundreds of millions of times. We proposed obtaining RNA samples from viruses, bacteria, and plants for the reverse transcription reactions with a subsequent amplification of cDNAs by the polymerase chain reaction using potent and nontoxic chaotropic agent ammonium trichloroacetate. The method works in the analytical and preparative range and can be useful in the case of extraordinary circumstances during mass infections. Potentially this method can be adapted for obtaining RNA samples ready for the RT-isothermal PCR in the field. [Display omitted] •Isolation of RNA with ammonium trichloroacetate/SDS from viruses, cells, and tissue is an environmentally friendly method.•This method is appropriate in extraordinary circumstances for the detection of RNA virus infections by RT-PCR.•Potentially the method can be automated and adapted for use in the field.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2021.114113