大鼠骨骼肌RNA提取工艺的优化

S. Felipe, Christina Pacheco, J. E. R. Martins, Raquel Martins de Freitas, Paulo Elesson Oliveira, S. V. D. Mendes, J. Alves, V. Ceccatto
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引用次数: 0

摘要

目的:本研究旨在建立和表征一种优化的方案构象,以从啮齿动物骨骼肌样本中获得足够的RNA质量,用于测序研究。研究地点和时间:体内实验和分析在ceear州立大学生物医学科学高级研究所生物化学和基因表达实验室(LABIEX)进行。在2017 - 2020之间。方法:选取23例雄性Wistar大鼠骨骼肌,特别是比目鱼肌。总RNA提取使用经典的TRIzol®方法和商业试剂盒,合并两者的步骤。生物分析仪平台毛细管电泳用于RNA质量评价。结果:(C)对适应方案的RNA浓度、RIN和28S/18S率进行分析,结果令人满意。28S/18S核糖体条带清晰,没有小的痕迹,表明RNA具有高完整性,没有基因组DNA的污染。结论:获得的RNA质量和完整性数据满足后路RNA测序的要求。
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Optimization of RNA Extraction Protocol for Rat Skeletal Muscle Samples
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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