Co-Extraction of DNA and RNA from Candida albicans Using a Chemical Method in Conjunction with Silicon Carbide with Few Cells
Authors:
- Elizabeth Cristina Vieira de FreitasEFElizabeth Cristina Vieira de Freitas
- Francisca Alves dos SantosFSFrancisca Alves dos Santos
- Maria Raíssa Vieira LopesMLMaria Raíssa Vieira Lopes
- Dárcio Luiz de Sousa JúniorDJDárcio Luiz de Sousa Júnior
- Tássia Thaís Al YafawiTYTássia Thaís Al Yafawi
- Ana Carolina Ferreira AraújoAAAna Carolina Ferreira Araújo
- Priscilla Ramos FreitasPFPriscilla Ramos Freitas
- Irwin Rose Alencar de MenezesIrwin Rose Alencar de Menezes
- Henrique Douglas Melo CoutinhoHCHenrique Douglas Melo Coutinho
- Maria Karollyna Do Nascimento Silva LeandroMLMaria Karollyna Do Nascimento Silva Leandro
Open Access
Abstract
Objective: The study aimed to optimize protocols for the joint extraction of deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) from 0.025 × 106 CFU of Candida albicans, targeting to overcome the challenges in the extraction of these genetic materials. Materials and methods: From this, treated silicon carbide (SiC) granules were added to fungal samples from methods 1, 2, and 3 obtained from aliquots of BHI or Sabouraud medium to cause cell lysis and enable the isolation of these macromolecules by phenol and chloroform. The concentration and integrity of the extracted nucleic acids were analyzed, respectively, by spectrophotometry using the A260/A280 ratios and 1% agarose gel electrophoresis. Results: Therefore, method 3 is the one that most comprises samples considered pure of both DNA and RNA, simultaneously. Furthermore, the presence of intact RNAs corresponding to the base pair size such as 5.8 S rRNA and tRNA was verified during electrophoresis, considering the particularities of RNA, which makes it very unstable and easily degraded. Conclusions: Thus, it results in a faster and simpler method in addition to obtain promising results using minimal amounts of biological sample and offering a valuable alternative for small laboratories to work with molecular biology.Keywords
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This publication has 18 references indexed in Scilit:
- Candida albicans - Biology, molecular characterization, pathogenicity, and advances in diagnosis and control – An updateMicrobial Pathogenesis, 2018
- Alternative methodology for extraction of high-quality DNA from ancient bones by demineralization without pulverizationForensic Science and Criminology, 2018
- Microbiological control of parenteral dosage formsBiomedical and Biopharmaceutical Research, 2016
- Isolamento de DNA genômico a partir de folhas secas de Erythrina crista-galli L., FABACEAE (Corticeira-do-banhado)Revista Thema, 2015
- Looking intoCandida albicansinfection, host response, and antifungal strategiesVirulence, 2015
- In vitro organogenesis of zucchini squash cv. CasertaAdvances in Rheumatology, 2012
- Método rápido de extração de DNA de bactériasSumma Phytopathologica, 2008
- Pesquisa em biologia molecular: como fazer?Revista Brasileira de Coloproctologia, 2006
- Current two‐dimensional electrophoresis technology for proteomicsProteomics, 2004
- Extração de DNA de materiais de arquivo e fontes escassas para utilização em reação de polimerização em cadeia (PCR)Revista Brasileira de Ginecologia e Obstetrícia, 2004