Hello, I am trying to optimize sonication for ChIP. I am working with aml cell lines viz. HL60 U937 and THP1. I have fixed the cells with 1% formaldehyde and stored at -80 degrees. There are two things that I struggle with:
1. Amount of time I have to keep cells in hypotonic buffer for cell lysis.
2. Ideal number of cycles to shear the chromatin.
I know the size should be below 500bp for ChIP. But it would be great if I could get some inputs if anyone is working with these cell lines and optimized ChIP for the same. Oh and yes. I sonicate 2.5 million cells
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