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urgent: need help with qPCR statistics


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#1 joy123

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Posted 26 April 2013 - 08:06 AM

Hi, I did qPCR on 16S gene to test the level of a bacteria from four groups of people.

I don't have standard curve when I did qPCR. But in each plate I included one standard bacteria strain gene (in triplicate). I want to ask how should I analyze the data. Should I simply compare Ct value by nonparametric analysis? Or shoud I do some others (like delta CT?)?

I see one paper compare gene difference in patients and healthy control by "fold change". But I don't know what they used to compare. Just compare Ct?

I need the results be analyzed by the end of the day. Would you please help me with that?

Thanks a lotu

#2 pcrman

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Posted 26 April 2013 - 08:30 AM

Hi Joy123,

what is your internal control - the control that tells you a difference in the Ct of bacteria 16S comes from a difference in infection or in unequal starting RNA? If you have included an internal control, then you use delta Ct to obtain means for each group of patients, and then use ANOVA to compare among groups.

#3 joy123

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Posted 26 April 2013 - 09:37 AM

Thanks for your reply! Unfortunately, my sample is a mixture of human cells and bacteria, and the cell munbers vary among people. So I don't have internal control. I loaded same amount of DNA (which has uncertain amount of human and bateria DNA) in the qPCR analysis, and compared the level of 16S gene to compare the amount of my interested bacteria in same amount of samples of different groups of people.




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