chip seq protocol - (Apr/26/2012 )
I would like to do Chip seq technique to my gene of interest. The problem I find many protocols about that and they use different lysis buffer like ( SDS lysis buffer,, or whole cell lysis buffer (abcam) ,, farnham cell lysis buffer)
In my experiment I will crosslink cells and carry on the Chip:
1- Does anyone have a protocol of chip seq with cancer cells like ( SW480, NT2)?
2- I am confused between using farnham lysis buffer or SDS lysis buffer whats the different ?
3- if I would like to work in only nuclei protien what lysis buffer and technique i shoud use?
I suggest you that do not waste your time with chip-seq.
Chep-exo is 30% better than that.
This company which is the owner of patent of chip-exo can do chip-exo for you with just 1400$.
this is the instruction that I have received from this company:
Babak from McGill
If you want yet to waste your time ok this is a sample I have:
Thank you for your replay.. and it is a good idea to use Chip-exo .. but for my project I have to stick with chip seq ..
I do not know if anyone can answer my questions above cause I need to know whats is the different:
Farnham lysis buffer uses NP40 as a detergent to lyse the cell membrane but it not the nuclear membrane. The nuclei are then subsequently transferred to an SDS nuclear lysis/sonication buffer to lyse the nuclei and release the chromatin.
If you use an SDS lysis buffer from the start you can perform all the lysis and sonication in the same buffer, but you may get interference or background from cytoplasmic proteins.
Here's some good information: