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Different MOI in comparison experiment

Viro

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#1 Mohamed 1984

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Posted 31 January 2014 - 09:17 AM

I infected two cells with IAV using two different MOI, one of them 1 and the other is 10. Now I wanted to compare the expression level of one miRNA in both  cells. I think this is not right to compare. Since I did the experiment, has anyone any suggestion to surmount or overcome this mistake without repeating the experiment.   any suggestion



#2 bob1

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Posted 31 January 2014 - 01:29 PM

Why is it not right - as far as I can see, all you are comparing is the difference between two different infection amounts, so there shouldn't be a problem.  The results can be normalized to a cellular gene that isn't affected by the infection.



#3 Mohamed 1984

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Posted 31 January 2014 - 06:51 PM

But the problem I think is that when u hit the cells with a high MOI, more cells died while still  a live in the other low MOI and this affect the expression of cell genes. Which to normalize to  a gene that not affected by infection ? like B actin For example ?



#4 bob1

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Posted 31 January 2014 - 11:50 PM

Yes, dying cellare a problem, however DNA and RNA are rapidly degraded during apoptosis.

 

As to the gene - check the literature.  many viruses alter actin expression.






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