Kocabas Arif Murat, Marin-Valencia Isaac
The Abimael Laboratory of Neurometabolism, Department of Neurology, Icahn School of Medicine at Mount Sinai, New York, NY, USA.
The Abimael Laboratory of Neurometabolism, Department of Neurology, Icahn School of Medicine at Mount Sinai, New York, NY, USA; Departments of Neuroscience, Genetics and Genomics Medicine, and Pediatrics Icahn School of Medicine at Mount Sinai, New York, NY, USA.
STAR Protoc. 2024 Dec 20;5(4):103495. doi: 10.1016/j.xpro.2024.103495. Epub 2024 Dec 12.
Isolating RNA from single nuclei is essential for single-cell gene expression analysis, yet obtaining high-quality RNA is challenging. We present a protocol to enhance RNA yield and quality from mouse brain nuclei. Key steps include brain dissection, thawing, homogenization, and centrifugation-based isolation. The protocol incorporates 3% glyoxal fixation for RNA preservation, followed by filtration, blocking, and fluorescence-activated sorting to ensure the extracted RNA meets quality and quantity standards for transcriptomic and qPCR analyses. For complete details on the use and execution of this protocol, please refer to Marin-Valencia, Kocabas et al..
从单细胞核中分离RNA对于单细胞基因表达分析至关重要，但获得高质量RNA具有挑战性。我们提出了一种提高从小鼠脑细胞核中提取RNA产量和质量的方案。关键步骤包括脑解剖、解冻、匀浆以及基于离心的分离。该方案采用3%乙二醛固定以保存RNA，随后进行过滤、封闭和荧光激活分选，以确保提取的RNA符合转录组学和qPCR分析的质量和数量标准。有关此方案的使用和执行的完整详细信息，请参考Marin-Valencia、Kocabas等人的研究。
