RNA extraction from the yeast candida parapsilosis sensu stricto using two commercially available silica column-based purification methods
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Date
2018
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Abstract
Good quality ribonucleic acid (RNA) needs to be obtained in order to study gene expression. Different RNA extraction methods have been described, but RNA quality and yield may vary among the different techniques and biological study species. To date, there is no standardized method for extraction and purification of RNA from Candida genus yeasts. The few available papers on the subject apply mainly to filamentous fungi and have produced poor results for extraction techniques based on manual methods. The aim of this study was therefore to compare two commercial RNA extraction and purification systems using silica columns (Qiagen and Zymo Research) with Candida parapsilosis sensu stricto as model organism. This yeast has been identified in recent papers as the second most frequently isolated Candida species in the oral cavity. In the past decade, it has been the object of increasing medical interest because it is one of the main causes of candidemia in both adults and preterm neonates. In view of this background, we consider the study of Candida parapsilosis sensu stricto transcriptome and its variations according to environmental changes to be a priority. In this experimental study, 19 fungal isolates were processed using Qiagen and 17 isolates using Zymo Research. The results suggest that Qiagen lysis buffer RLT is essential for obtaining better quality RNA product.
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Keywords
RNA extraction, RNA purification, Candida parapsilosis sensu stricto, Silica columns