Análise comparativa de métodos de extração visando a obtenção de RNA de qualidade na cultura da mandioca

Abstract

Cassava (Manihot esculenta Crantz) is considered one of the main agricultural crops, not only in Brazilian territory but also in several other countries. To understand the molecular processes that regulate the development and resistance of cassava to diseases, for example, the use of advanced molecular tools is essential. RNA extraction is a fundamental step in molecular studies. However, the removal process is complex and requires care to avoid degradation of the RNA and ensure sample purity. The objective of this work was to perform a comparative analysis of three RNA isolation methods and identify the most appropriate protocol for extract quality RNA from three cassava tissues: tuberous root, stem and leaf. The CTAB methods (two precipitation way), TRIzol and Acid Phenol were compared. Of the three extraction methods tested in this work, the CTAB method, with optimizations using lithium chloride precipitation, allowed the isolation of high-quality RNA from root, stem and leaf with the suggested levels of quantity and purity and it was sufficient for the development of molecular assays.