Optimization of RNA Extraction Protocol for Rat Skeletal Muscle Samples
Stela Mirla da Silva Felipe
Instituto Superior de Ciências Biomédicas, ISCB – Universidade Estadual do Ceará, Fortaleza, Brazil.
Christina Pacheco
Núcleo de Medicina Tropical, Universidade de Brasília, Brasília, Distrito Federal, Brazil.
Jonathan Elias Rodrigues Martins
Instituto Superior de Ciências Biomédicas, ISCB – Universidade Estadual do Ceará, Fortaleza, Brazil.
Raquel Martins de Freitas *
Instituto Superior de Ciências Biomédicas, ISCB – Universidade Estadual do Ceará, Fortaleza, Brazil.
Paulo Elesson Guimarães de Oliveira
Instituto Superior de Ciências Biomédicas, ISCB – Universidade Estadual do Ceará, Fortaleza, Brazil.
Sávio Victor Diógenes Mendes
Instituto Superior de Ciências Biomédicas, ISCB – Universidade Estadual do Ceará, Fortaleza, Brazil.
Juliana Osório Alves
Centro Universitário Estácio - FIC - Fortaleza, Ceará, Brazil.
Vânia Marilande Ceccatto
Instituto Superior de Ciências Biomédicas, ISCB – Universidade Estadual do Ceará, Fortaleza, Brazil.
*Author to whom correspondence should be addressed.
Abstract
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.
Keywords: Sequencing, RNA extraction, soleus muscle
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References
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