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; Hippenmeyer, Simon; Role of Eed in neural stem cell lineage progression; Molecular Mechanisms of Neural Stem Cell Lineage Progression; Mapping Cell-Type Specificity of the Genomic Imprintome in the Brain; Molecular Mechanisms of Cerebral Cortex Development; Principles of Neural Stem Cell Lineage Progression in Cerebral Cortex Development (STAR Protocols, 2020)
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).
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Rights: https://creativecommons.org/licenses/by-nc-nd/4.0/
Keywords: article