Staining for Cell Cycle Analysis How does propidium iodide measure the cell cycl
ID: 3165081 • Letter: S
Question
Staining for Cell Cycle Analysis
How does propidium iodide measure the cell cycle?
Why do you have to get rid of RNA?
Why are the components of the staining solution dissolved in PBS rather than in water?
Why cant this assay distinguish between G2 and M?
QUESTION 1 How does propidium iodide measure the cell cycle? Its fluorescence is proportional to the amount of DNA It's fluorescence is proportional to the amount of protein O rs tracks te cell cycle protein expression O its fuorescence is proportional to the amount of ipid QUESTION 2 Why do you have to get rid of RNA? Propidium iodide stains both DNA and RNA RNA would gum up the instrumentation O Cells won't cycle if RNA is present RNA causes cells to dye QUESTION 3 Why are the components of the staining solution dissolved in PBS rather than in water? Propid un lodide cant be di Water would cause the cells to lyse QUESTION 4 Why can't this assay distinguish between G2 and M7 O There is too much RNA at that time O They have the same proteins It is not sensitive enough They have the same amount of DNA QUESTION 5 Are the cells you are working with in this assay synchronous)Explanation / Answer
Question 1) Propidium iodide is used to measure the cell cycle because it stains DNA quantitatively. Firstly,cells are permeabilised and treated with this fluorescent dye,the fluorescent intensity is directly proportional to the amount of DNA present. Example-if the cell is in S phase,then the intensity of fluorescence is double than that of G0 or G1 phase because DNA is replicated in S phase.
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