Hello all,
I have been doing qChIP successfully for a while now, and I am trying to make the venture into ChIP-seq. My problem is that I have very small samples (vertebrate embryos) and after I pool and concentrate and do my magic ChIP dance, I get yields of about 2-8ng per ChIP (10% Input: 25-77ng).
The issue I'm having is that those concentrations are sort of in the middle of low-but-usable (~10ng) and the super low concentrations people like to do amplification (LinDA, WGA) or modified library prep (eg. Bernstein's Nano-ChIP-seq).
I feel like there's a gray area in this low (but not micro-low) levels where I'm not sure the best way to proceed.
Also, I remember reading somewhere that some people would spike their IP with input DNA to get the concentrations up. This seems like it would have massive affects on quantification, right?
Thanks for your help. This forum is seriously the greatest.
Bob
I have been doing qChIP successfully for a while now, and I am trying to make the venture into ChIP-seq. My problem is that I have very small samples (vertebrate embryos) and after I pool and concentrate and do my magic ChIP dance, I get yields of about 2-8ng per ChIP (10% Input: 25-77ng).
The issue I'm having is that those concentrations are sort of in the middle of low-but-usable (~10ng) and the super low concentrations people like to do amplification (LinDA, WGA) or modified library prep (eg. Bernstein's Nano-ChIP-seq).
I feel like there's a gray area in this low (but not micro-low) levels where I'm not sure the best way to proceed.
Also, I remember reading somewhere that some people would spike their IP with input DNA to get the concentrations up. This seems like it would have massive affects on quantification, right?
Thanks for your help. This forum is seriously the greatest.
Bob
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