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
Preprint
- 13 September 2024
- preprint
- Published by MDPI AG in Preprints.org
Abstract
The study aimed to optimize protocols for the joint extraction of deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) from 0.025 x 106 CFU of Candida albicans, aiming to overcome challenges in the extraction of these genetic materials. 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 nucleic acids were obtained, respectively, by spectrophotometry using A260/A280 ratios and 1% agarose gel electrophoresis. 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, taking into account the particularities of RNA, which makes it very unstable and easily degraded. Therefore, it results in a faster and simpler method in addition to obtaining promising results using minimal amounts of biological sample, thus offering a valuable alternative for small laboratories working with molecular biology.Keywords
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