ChIP assay! - (Mar/06/2007 )
Hi everyone,
I'm trying to perform the ChIP assay, and it seems that I could be successful. However, I dun know how to quantify the results. According to the mannual, the manufacturer suggested that after isolating chromosome DNA, you should use the same volumn of DNA for PCR. But, in my opinion, when you lyse the cells, collect the cells, ....you can not guarantee that the yield for all samples are the same. I mean that you can not be sure that in the same volumn, all of your samples contain the same DNA amount.
It's my problem, and to solve this, I determine the DNA concentration of the Input (before IP) and apply the ratio (between the control and stimulated Input samples) to the samples that were performed IP with specific antibody. I wondered if this is true. Could you please help me!
Thank you so much!
I'm trying to perform the ChIP assay, and it seems that I could be successful. However, I dun know how to quantify the results. According to the mannual, the manufacturer suggested that after isolating chromosome DNA, you should use the same volumn of DNA for PCR. But, in my opinion, when you lyse the cells, collect the cells, ....you can not guarantee that the yield for all samples are the same. I mean that you can not be sure that in the same volumn, all of your samples contain the same DNA amount.
It's my problem, and to solve this, I determine the DNA concentration of the Input (before IP) and apply the ratio (between the control and stimulated Input samples) to the samples that were performed IP with specific antibody. I wondered if this is true. Could you please help me!
Thank you so much!
hi there,
I think you should use the same DNA amount of input to perform chip assay. But i am not sure how to adjust volume of the antibody IP samples?
Thanks