Protocol for enhancing RNA yield and quality from single nuclei isolated from mouse brain tissue
Summary: 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 centrifugat...
Saved in:
| Main Authors: | , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2024-12-01
|
| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724006609 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850244903301283840 |
|---|---|
| author | Arif Murat Kocabas Isaac Marin-Valencia |
| author_facet | Arif Murat Kocabas Isaac Marin-Valencia |
| author_sort | Arif Murat Kocabas |
| collection | DOAJ |
| description | Summary: 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.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-9a74e9de869b42dba7a9516b0bb701e5 |
| institution | OA Journals |
| issn | 2666-1667 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-9a74e9de869b42dba7a9516b0bb701e52025-08-20T01:59:35ZengElsevierSTAR Protocols2666-16672024-12-015410349510.1016/j.xpro.2024.103495Protocol for enhancing RNA yield and quality from single nuclei isolated from mouse brain tissueArif Murat Kocabas0Isaac Marin-Valencia1The Abimael Laboratory of Neurometabolism, Department of Neurology, Icahn School of Medicine at Mount Sinai, New York, NY, USA; Corresponding authorThe 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; Corresponding authorSummary: 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.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166724006609MetabolismNeuroscience |
| spellingShingle | Arif Murat Kocabas Isaac Marin-Valencia Protocol for enhancing RNA yield and quality from single nuclei isolated from mouse brain tissue STAR Protocols Metabolism Neuroscience |
| title | Protocol for enhancing RNA yield and quality from single nuclei isolated from mouse brain tissue |
| title_full | Protocol for enhancing RNA yield and quality from single nuclei isolated from mouse brain tissue |
| title_fullStr | Protocol for enhancing RNA yield and quality from single nuclei isolated from mouse brain tissue |
| title_full_unstemmed | Protocol for enhancing RNA yield and quality from single nuclei isolated from mouse brain tissue |
| title_short | Protocol for enhancing RNA yield and quality from single nuclei isolated from mouse brain tissue |
| title_sort | protocol for enhancing rna yield and quality from single nuclei isolated from mouse brain tissue |
| topic | Metabolism Neuroscience |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724006609 |
| work_keys_str_mv | AT arifmuratkocabas protocolforenhancingrnayieldandqualityfromsinglenucleiisolatedfrommousebraintissue AT isaacmarinvalencia protocolforenhancingrnayieldandqualityfromsinglenucleiisolatedfrommousebraintissue |