Harvesting Cultured Dopaminergic Neurons for RNA Sequencing
7:04
Generation, Purification and Amplification of First-strand cDNA
9:13
Results: RNA-seq Detection of Expressed Transcripts in Single Dopaminergic Neurons
9:39
Conclusion
Transcripción
The overall goal of this procedure is to reliably identify living dopaminergic neurons in midbrain cultures using tyrosine hydroxlase driven eGFP expression, single cell harvesting, and RNA sequencing library generation. This method can help answe
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In Parkinson's Disease (PD), Substantia Nigra (SNc) dopaminergic neurons degenerate, leading to motor dysfunction. Here we report a protocol for culturing ventral midbrain neurons from a mouse expressing eGFP driven by a Tyrosine Hydroxylase (TH) promoter sequence, harvesting individual fluorescent neurons from the cultures, and measuring their transcriptome using RNA-seq.