I would like to perform ChIP seq for Histone modifications on primary cells.The number of cells are below 10,000. I would appreciate help from someone who has experience working with such low number ChIP protocols. I have read some papers about low ChIP though. Any suggestions or tricks are welcome. Thank you
Hi!
Chip using small numbers of cells is generally tricky. You could try those protocols, mainly by Dahl I assume. Uncommon modifications are going to be tough though, at low cell counts. I have had success with a slightly modified version of a previously published protocol. You can find the paper on my profile (eldholm 2014 ijc), in which the original protocol is also referenced. And stay away from Santa Cruz abs unless they have been thoroughly verified. They very often suck.
Cheers,
Vegard
Hi,
In your case I would recommend to follow protocols in which the PCR cycle is determi
Dear Pavan,
Thanks a lot
I already had look at iChIP protocol earlier and trying to work on it.Seems to be very helpful.Lets see,If that works..Still discussing with sequencing facility for adapter ligation etc.Will update you ..
Could try this native ChIP approach: http://www.nature.com/ncomms/2015/150121/ncomms7033/full/ncomms7033.html
My lab developed this protocol, we have used it to do ChIP-PCR from as few as 50 cells.
http://www.slideshare.net/cwynder/small-volume-histone-prep-es-cs
We also published it as part of a larger methods collection for defining HDM activity in Stem cells
http://www.springerlink.com/content/rmk2r0514k3l3l43/#section=993322&page=1
We have a high sensitivity ChIP kit from 2,000 cells input:
http://www.epigentek.com/catalog/chromaflash-high-sensitivity-chip-kit-p-3620.html
We also have an all-in-one ChIP + Illumina library prep kit for ChIP-seq:
http://www.epigentek.com/catalog/epinext-chip-seq-high-sensitivity-kit-illumina-p-3650.html
Good luck
Dear Nick,
Thanks a lot for the informationa nd I would look into more details in the brochure.
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