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Next-Generation Sequencing Application: A Systematic Approach for High-Quality RNA Isolation from Skeletal Muscles.
Hernandez-Morato, Ignacio; Kemfack, Angela M.
Afiliación
  • Hernandez-Morato I; Department of Anatomy and Embryology, School of Medicine, Complutense University of Madrid, Madrid, Spain. ighernan@ucm.es.
  • Kemfack AM; The Center for Voice and Swallowing, Department of Otolaryngology-Head & Neck Surgery, Columbia University Irving Medical Center/New York Presbyterian, New York, NY, USA.
Methods Mol Biol ; 2822: 13-24, 2024.
Article en En | MEDLINE | ID: mdl-38907908
ABSTRACT
RNA extraction and analyses from tissues using bulk RNA-Sequencing (RNA-Seq) provide a more accurate picture of the gene expression compared to other molecular biology techniques for RNA quantification. Challenges associated with high-quality RNA extraction from skeletal muscles require a modification of standard protocols. Here, we describe a procedure for high-quality RNA isolation from intrinsic laryngeal muscles transferable to skeletal muscles with comparable technical and biological difficulties. Standard protocols for RNA isolation were optimized by maximizing the pooling strategy, determining the sample weight, applying cryogenic muscle disruption, and incorporating RNase-inhibiting reagents during the tissue preparation steps.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: ARN / Análisis de Secuencia de ARN / Músculo Esquelético / Secuenciación de Nucleótidos de Alto Rendimiento Límite: Animals Idioma: En Revista: Methods Mol Biol Asunto de la revista: BIOLOGIA MOLECULAR Año: 2024 Tipo del documento: Article País de afiliación: España Pais de publicación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: ARN / Análisis de Secuencia de ARN / Músculo Esquelético / Secuenciación de Nucleótidos de Alto Rendimiento Límite: Animals Idioma: En Revista: Methods Mol Biol Asunto de la revista: BIOLOGIA MOLECULAR Año: 2024 Tipo del documento: Article País de afiliación: España Pais de publicación: Estados Unidos