Real-time quantification in Chromatin IP - (Jan/11/2005 )
I would like to compare precipitated target DNA in Chromatin Immuno Precipitation by real-time quantitative PCR. How should I compare Ct values of untreated, treated (no antibody) and treated (with antibody) sets taking care of Inputs. How should I plot? Could you please explain? I would appreciate the suggestion of any paper as reference too.
-utpal-
One of the things you would need to do is accurately quantitate your DNA.
We do it by picogreen/Quant-It assays from Invitrogen, we then perform a q-PCR with the specific primers of interest adding equivalent amounts of DNA from each sample.
Good luck.
Nick
-methylnick-
QUOTE (methylnick @ Jan 29 2007, 06:38 PM)
One of the things you would need to do is accurately quantitate your DNA.
We do it by picogreen/Quant-It assays from Invitrogen, we then perform a q-PCR with the specific primers of interest adding equivalent amounts of DNA from each sample.
Good luck.
Nick
We do it by picogreen/Quant-It assays from Invitrogen, we then perform a q-PCR with the specific primers of interest adding equivalent amounts of DNA from each sample.
Good luck.
Nick
Can anyone on this forum has link to excel work sheet template for analyzing CHIP data with real time pcr. As I understand from ct values (of vector and and transfceted cell IP) first we deduct from input and then to find the fold change in transfcted cells we deuct their ct from vector cells followed by calculating power.
Please correct me if I am wrong please.
Thanks.
-honeyhappy-