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