Optimization of RNA Extraction Protocol for Rat Skeletal Muscle Samples
Stela Mirla Da Silva Felipe, Christina Pacheco, Jonathan Elias Rodrigues Martins, Raquel Martins De Freitas, Paulo Elesson Guimarães de Oliveira, Sávio Victor Diógenes Mendes, Juliana Osório Alves, Vânia Marilande Ceccatto
Journal of Applied Life Sciences International · pp. 10–16 · Published 7 Jan 2023
10.9734/jalsi/2023/v26i1592Abstract
Aims: The present study aimed to stablish and characterize an optimized protocol conformation to obtain adequate RNA quality from rodents skeletal muscle samples for sequencing studies. Place and Duration of Study: The in vivo experiments and analyses were performed in the Laboratory of Biochemistry and Gene Expression – LABIEX of the Superior Institute of Biomedical Science – ISCB from the State University of Ceará - UECE. Between 2017-2020. Methodology: Were used 23 samples from male Wistar rat skeletal muscle, specifically from soleus muscle. Total RNA extraction was performed using the classic TRIzol® method and commercial kit, merging steps from both. Capillary electrophoresis in the Bioanalyzer platform was used for RNA quality evaluation. Results: (C) Analyzes of adapted protocol RNA concentration, RIN and rate 28S/18S showed satisfactory results. 28S/18S Ribosomal bands appear well defined, without small traces, which indicates RNA with high integrity and without contamination of genomic DNA. Conclusion: Obtained RNA quality and integrity data satisfied the exigencies for posterior RNA-seq.
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