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Isolation and culturing of olfactory sensory neurons

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Original posting as formulated by the Student:

I am writing a research proposal in which I am attempted to describe an experimental procedure to isolate olfactory sensory neurons (OSNs) and analyze/compare the amino acid sequences of olfactory receptors expressed by individual OSNs. To do this, I am proposing isolating individual OSNs of interest, extracting RNA, and conducting reverse transcription to generate cDNA for PCR and sequencing.

The problem I am having is I am not sure if a single OSN is adequate for RNA extraction and reverse transcription. If not, I would need to describe a method for isolating and culturing individual OSNs to produce enough clones for RNA extraction. If a single cell is not enough, what is a way that clones could be generated from the single OSN?

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Solution Summary

The procedure describes isolation and culturing olfactory sensory neurons (OSN) from the mouse brain.

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Ideally, one would want to generate a culture of identical cells from a single cell. This task might be challenging because single neurons normally do not survive in culture. Therefore, the procedure would require isolating a population of neurons and culturing them in optimal conditions.

A solution provides the protocol for isolating and culturing OSNs. The protocol is based on a published research article, where the procedure was used. The protocol is also attached as a word document.

Preparation of an olfactory sensory neurons (OSN) primary cell culture from the mouse brain.

The OSN ...

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