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Generation and isolation of single cells from mouse brain with mosaic analysis with double markers-induced uniparental chromosome disomy
Mosaic analysis with double markers (MADM) technology enables concomitant fluorescent cell labeling and induction of uniparental chromosome disomy (UPD) with single-cell resolution. In UPD, imprinted genes are either overexpressed 2-fold or are not expressed. Here, the MADM platform is utilized to p...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7757670/ https://www.ncbi.nlm.nih.gov/pubmed/33377108 http://dx.doi.org/10.1016/j.xpro.2020.100215 |
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author | Laukoter, Susanne Amberg, Nicole Pauler, Florian M. Hippenmeyer, Simon |
author_facet | Laukoter, Susanne Amberg, Nicole Pauler, Florian M. Hippenmeyer, Simon |
author_sort | Laukoter, Susanne |
collection | PubMed |
description | Mosaic analysis with double markers (MADM) technology enables concomitant fluorescent cell labeling and induction of uniparental chromosome disomy (UPD) with single-cell resolution. In UPD, imprinted genes are either overexpressed 2-fold or are not expressed. Here, the MADM platform is utilized to probe imprinting phenotypes at the transcriptional level. This protocol highlights major steps for the generation and isolation of projection neurons and astrocytes with MADM-induced UPD from mouse cerebral cortex for downstream single-cell and low-input sample RNA-sequencing experiments. For complete details on the use and execution of this protocol, please refer to Laukoter et al. (2020b). |
format | Online Article Text |
id | pubmed-7757670 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-77576702020-12-28 Generation and isolation of single cells from mouse brain with mosaic analysis with double markers-induced uniparental chromosome disomy Laukoter, Susanne Amberg, Nicole Pauler, Florian M. Hippenmeyer, Simon STAR Protoc Protocol Mosaic analysis with double markers (MADM) technology enables concomitant fluorescent cell labeling and induction of uniparental chromosome disomy (UPD) with single-cell resolution. In UPD, imprinted genes are either overexpressed 2-fold or are not expressed. Here, the MADM platform is utilized to probe imprinting phenotypes at the transcriptional level. This protocol highlights major steps for the generation and isolation of projection neurons and astrocytes with MADM-induced UPD from mouse cerebral cortex for downstream single-cell and low-input sample RNA-sequencing experiments. For complete details on the use and execution of this protocol, please refer to Laukoter et al. (2020b). Elsevier 2020-12-16 /pmc/articles/PMC7757670/ /pubmed/33377108 http://dx.doi.org/10.1016/j.xpro.2020.100215 Text en © 2020 The Author(s) http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Protocol Laukoter, Susanne Amberg, Nicole Pauler, Florian M. Hippenmeyer, Simon Generation and isolation of single cells from mouse brain with mosaic analysis with double markers-induced uniparental chromosome disomy |
title | Generation and isolation of single cells from mouse brain with mosaic analysis with double markers-induced uniparental chromosome disomy |
title_full | Generation and isolation of single cells from mouse brain with mosaic analysis with double markers-induced uniparental chromosome disomy |
title_fullStr | Generation and isolation of single cells from mouse brain with mosaic analysis with double markers-induced uniparental chromosome disomy |
title_full_unstemmed | Generation and isolation of single cells from mouse brain with mosaic analysis with double markers-induced uniparental chromosome disomy |
title_short | Generation and isolation of single cells from mouse brain with mosaic analysis with double markers-induced uniparental chromosome disomy |
title_sort | generation and isolation of single cells from mouse brain with mosaic analysis with double markers-induced uniparental chromosome disomy |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7757670/ https://www.ncbi.nlm.nih.gov/pubmed/33377108 http://dx.doi.org/10.1016/j.xpro.2020.100215 |
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