Protocol for sorting cells from mouse brains labeled with mosaic analysis with double markers by flow cytometry
Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved..
Mosaic analysis with double markers (MADM) technology enables the generation of genetic mosaic tissue in mice and high-resolution phenotyping at the individual cell level. Here, we present a protocol for isolating MADM-labeled cells with high yield for downstream molecular analyses using fluorescence-activated cell sorting (FACS). We describe steps for generating MADM-labeled mice, perfusion, single-cell suspension, and debris removal. We then detail procedures for cell sorting by FACS and downstream analysis. This protocol is suitable for embryonic to adult mice. For complete details on the use and execution of this protocol, please refer to Contreras et al. (2021).1.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2024 |
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| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:5 |
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| Enthalten in: |
STAR protocols - 5(2024), 1 vom: 15. März, Seite 102771 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Amberg, Nicole [VerfasserIn] |
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| Links: |
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| Themen: |
Developmental biology |
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| Anmerkungen: |
Date Completed 20.03.2024 Date Revised 20.03.2024 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1016/j.xpro.2023.102771 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| 520 | |a Mosaic analysis with double markers (MADM) technology enables the generation of genetic mosaic tissue in mice and high-resolution phenotyping at the individual cell level. Here, we present a protocol for isolating MADM-labeled cells with high yield for downstream molecular analyses using fluorescence-activated cell sorting (FACS). We describe steps for generating MADM-labeled mice, perfusion, single-cell suspension, and debris removal. We then detail procedures for cell sorting by FACS and downstream analysis. This protocol is suitable for embryonic to adult mice. For complete details on the use and execution of this protocol, please refer to Contreras et al. (2021).1 | ||
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| 700 | 1 | |a Hippenmeyer, Simon |e verfasserin |4 aut | |
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