Department of Anatomy and Embryology, School of Medicine, Complutense University of Madrid, Madrid, Spain.
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. 2024;2822:13-24. doi: 10.1007/978-1-0716-3918-4_2.
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.
使用批量 RNA 测序（RNA-Seq）从组织中提取和分析 RNA 与其他 RNA 定量的分子生物学技术相比，能更准确地反映基因表达情况。从骨骼肌中提取高质量 RNA 存在一些挑战，这需要对标准方案进行修改。在此，我们描述了一种从内在喉肌中提取高质量 RNA 的方法，该方法可转移到具有类似技术和生物学困难的骨骼肌。通过最大化混合策略、确定样品重量、采用低温肌肉破碎以及在组织准备步骤中加入 RNA 酶抑制剂等方式对 RNA 分离的标准方案进行了优化。
