RNA Extraction from the Yeast Candida parapsilosis Sensu Stricto Using Two Commercial Methods Based on Purification by Silica Columns

Abstract

There is no standardized methodology to date for extraction and purification of RNA from yeasts of the genus Candida. The few papers on the subject apply mainly to filamentous fungi, and have provided poor results for extraction techniques based on manual methods. The aim of this study was therefore to compare two commercial systems, Qiagen and Zymo Research, for extraction and purification of RNA with silica columns. The main differences between the two systems are the homogenization system used for cell suspension and cost. Material and Methods The Qiagen system was selected for its lysis buffer RLT, which has high concentrations of guanidine isothiocyanate and lacks phenol in its composition. As far as we know, there is no other system on the market with similar composition. The Zymo-research system was selected due to its lysis buffer based on a combination of phenol and guanidine, with the commercial name of Tri-reagent, which is equivalent to TRIzol®. These are two ideal systems to be compared in terms of their efficiency in obtaining good quantity and quality of RNA. A retrospective, cross-sectional, comparative basic research study was designed to compare two commercial systems for extraction and purification of RNA with silica columns regarding their ability to provide more and better quality biomolecules from the fungal cell biomass of the yeast Candida parapsilosis sensu stricto. A collection of 36 isolates was used, which had been characterized as Candida parapsilosis sensu stricto in a previous study using molecular methods. The strains were assigned randomly for processing with the Qiagen system (n=19) or Zymo Research system (n=17).