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ChIP/qPCR: IgG control, dilution of samples and enrichement


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#1 bioche82

bioche82

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Posted 11 September 2010 - 08:31 AM

Hello guys,

I have a few questions about ChIPs and quantitative real-time PCR.
When performing RT-PCR on ChIP samples, why do I have to dilute my samples 1/5? Is it because elements in the samples might affect the amplification and why would that be?

My other question is why exactly is the IgG control for and what could explain that I have a significant amplification for my IgG samples?

Finally, what could explain that I obtain no amplification at all with my qPCRs when I expected a binding of my transcription factor to my target DNA sequence?



Thank you very much in advance.




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