S. Felipe, Christina Pacheco, J. E. R. Martins, Raquel Martins de Freitas, Paulo Elesson Oliveira, S. V. D. Mendes, J. Alves, V. Ceccatto
{"title":"大鼠骨骼肌RNA提取工艺的优化","authors":"S. Felipe, Christina Pacheco, J. E. R. Martins, Raquel Martins de Freitas, Paulo Elesson Oliveira, S. V. D. Mendes, J. Alves, V. Ceccatto","doi":"10.9734/jalsi/2023/v26i1592","DOIUrl":null,"url":null,"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. \nPlace 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. \nMethodology: 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. \nResults: (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. \nConclusion: Obtained RNA quality and integrity data satisfied the exigencies for posterior RNA-seq.","PeriodicalId":14990,"journal":{"name":"Journal of Applied Life Sciences International","volume":"2 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2023-01-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Optimization of RNA Extraction Protocol for Rat Skeletal Muscle Samples\",\"authors\":\"S. Felipe, Christina Pacheco, J. E. R. Martins, Raquel Martins de Freitas, Paulo Elesson Oliveira, S. V. D. Mendes, J. Alves, V. Ceccatto\",\"doi\":\"10.9734/jalsi/2023/v26i1592\",\"DOIUrl\":null,\"url\":null,\"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. \\nPlace 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. \\nMethodology: 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. \\nResults: (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. \\nConclusion: Obtained RNA quality and integrity data satisfied the exigencies for posterior RNA-seq.\",\"PeriodicalId\":14990,\"journal\":{\"name\":\"Journal of Applied Life Sciences International\",\"volume\":\"2 1\",\"pages\":\"\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2023-01-07\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Applied Life Sciences International\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.9734/jalsi/2023/v26i1592\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Applied Life Sciences International","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.9734/jalsi/2023/v26i1592","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
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.