tm1987

Hi all,

I'm currently working with ChIP in my lab and my PI wants me to figure out a way to quantify the amount of chromatin I get from each tissue sample (mouse brain). I know you can quantify using a DNA concentration assay, but my PI wants me to try out a protein assay because it should be easier(?) Anyone heard of this?

Hi all,

I was wondering if anyone has a protocol for isolating nuclei from mouse brain tissue. I'm going to be using the nuclei for ChIP assay. I'm having trouble with the protocol I'm using now, as the nuclei are clumping together and it is very difficult to count them. Thanks.

Hi everyone

I'm having trouble with my nuclei clumping together, which makes it difficult for me to count them under the microscope. The protocol we use has us place the thawed tissue sample in PBS+PMSF and then fix the tissue in formaldehyde. After stopping the fixation with glycerine, we centrifuge the sample, remove the supernatant, and then resuspend the pellet in PBS+PMSF. From there we homogenize the tissue using needle and syringe. To count the nuclei, we add trypan blue to an aliquot of the sample and count the nuclei. Does anyone have an idea of what could be causing the nuclei to look like this? I'm not sure if the nuclei are just clumped together or if they're actually broken. Thanks in advance.