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