You have just read a paper that the addition of caffeine to cells disrupts the D

Question

You have just read a paper that the addition of caffeine to cells disrupts the DNA damage checkpoint and you would like to understand why. By searching for proteins that bind to caffeine, you isolate a protein, Caf1. You believe that Caf1 is involved in the DNA damage checkpoint because when you delete Caf1 from yeast cells and add a drug that causes DNA damage, the cells fail to arrest in the cell cycle. You give your Caf1-deleted cells to an undergraduate in the laboratory and ask him to take care of the strain for you while you take the weekend off to celebrate your finding. Upon your return, you find the poor undergraduate in tears. He explains that he thinks that he messed up the Caf1 mutant strain while you were gone because after growing Caf1 and wild-type cells in rich media and examining them during mitosis, Caf1 mutants looked the same as wild-type cells. Are you concerned about the undergraduate student’s findings? Why

Explanation / Answer

We should not concern with the silly mistake done by the undergrad student. Even if he messed up with the Caf1 mutant strain, we can find out which one is mutant and which one is the wildtype strain.

1- If we have antibody against Caf1 protein then it will be very easy for to find out mutant strain. We just need to isolate the protein from both the strain and run the Western blot the check which strain is giving us no band or which strain is giving us a band upon probing the blot with an anti-Caf1 antibody

2- If we don't have antibody against the Caf1 gene then alternatively we can isolate the mRNA from both the strain and run the qPCR to detect the presence or absence of any band with the Caf1 specific primers.

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