I am trying to make a ChIP, following by qPCR not by ChIPseq, from 10.000 facsed nuclei as starting material. Could someone suggest to me a protocol that could work with such a low amount of starting material? I look for bibliography but I have only found protocol for the equivalent amount of chromatin of 10.000 cells/ IP; in my case will be 2000 or less/IP.
I wili be very grateful if someone could help me.
Are you trying to assay histone modifications? With TF and cofactors it will be very challenging with such a low number of cells. Here are few links to published papers that have done ChIP with low numbers of cells:
doi: 10.1038/nmeth.3488
doi: http://dx.doi.org/10.1016/j.celrep.2015.10.004
doi: 10.1038/nmeth.3542
doi: 10.1038/ncomms7033
doi: 10.1126/science.1256271
doi: 10.1186/1471-2164-14-232
doi: 10.1038/nmeth.1626
doi: 10.1038/nbt.3383
Most of them are rather complex and do sequencing in the end but this can be easily replaced by qPCR.
Good luck!
Dear Jessica,
This looks like a perfect match with our True MicroChIP kit. It allows ChIP-seq from 10.000 cells.
Please let me know if you want to know more.
https://www.diagenode.com/p/true-microchip-kit-x16-16-rxns
You could incorporate carriers such as mRNA and Histone 2 to the lysate to increase the ChIP efficiency.
Here is the paper:
"A carrier-assisted ChIP-seq method for estrogen receptor-chromatin interactions from breast cancer core needle biopsy samples". BMC Genomics (2013).14:232
- Badges
- Science topic