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Protocol for sorting cells from mouse brains labeled with mosaic analysis with double markers by flow cytometry.

ABSTRACT: 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.

SUBMITTER: Amberg N

PROVIDER: S-EPMC10755489 | biostudies-literature | 2023 Dec

REPOSITORIES: biostudies-literature

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Protocol for sorting cells from mouse brains labeled with mosaic analysis with double markers by flow cytometry.

Amberg Nicole N Cheung Giselle G Hippenmeyer Simon S

STAR protocols 20231208 1

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 downs ...[more]

PMID: 38070137

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Protocol for sorting cells from mouse brains labeled with mosaic analysis with double markers by flow cytometry.

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Protocol for sorting cells from mouse brains labeled with mosaic analysis with double markers by flow cytometry.

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