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