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

Rodríguez Ml, Rosa Ac, Jewtuchowicz Vm
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Abstract

Good quality 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 or in-house IVD 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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以二氧化硅柱纯化为基础的两种商业方法提取严格假丝酵母RNA
为了研究基因表达,需要获得高质量的RNA。不同的RNA提取方法已经被描述,但RNA的质量和产量可能会因不同的技术和生物研究物种而异。迄今为止,还没有从念珠菌属酵母中提取和纯化RNA的标准化方法。关于该主题的少数可用论文主要适用于丝状真菌,并且基于手工或内部IVD方法的提取技术产生了较差的结果。因此,本研究的目的是比较使用二氧化硅柱(Qiagen和Zymo Research)的两种商用RNA提取和纯化系统,并将严格意义上的假丝酵母菌作为模式生物。这种酵母菌在最近的论文中被确定为口腔中第二常见的分离念珠菌。在过去的十年中,它一直是医学界越来越关注的对象,因为它是成人和早产儿念珠菌病的主要原因之一。鉴于这一背景,我们认为研究严格意义上的假丝酵母转录组及其随环境变化的变异是当务之急。在本实验中,19株真菌采用Qiagen法处理,17株真菌采用zimo Research法处理。结果表明,Qiagen裂解缓冲液RLT是获得高质量RNA产品的关键。
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