You are working with wild-type cells that have a 16h doubling time. BrdU is a mo

Question

You are working with wild-type cells that have a 16h doubling time. BrdU is a molecule that can be used to measure the amount of DNA inside cells. When added to cell culture, cells will incorporate BrdU into DNA strands during replication as if it were the DNA base thymidine (T) Your general experiment for using BrdU is: 1. Add BrdU to the media and let the cells continue to grow for 16h 2. Harvest cells and label them with a fluorescent antibody against Brdu 3. Measure fluorescence intensity in cells which is proportional to total DNA content 4. Plot a histogram which shows the number of cells with a certain fluorescence intensity

Explanation / Answer

B.)

Taxol is a drug which stabilizes the microtubules. As a result of which mitosis wont occur. All the cell will get arrested in the metaphase stage of mitosis. So first of all, the cells are in the 4n stage; second of all, after certain period of time all the cells from G1 phase moves to the M phase and get arrested. This in turn results in the shifting of graph to the right. We could observe only one peak in this graph, this is because all the cells are arrested in mitosis (with 4n DNA content) and also you could not find anymore cells in the G1 phase. So the peak associated with G1 phase (2n) falls and the peak associated with 4n chromosome rises. Because increase in DNA content will rise the peak and decrease in DNA content will make the peak to fall.

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