qPCR on chIP product - (Jul/03/2010 )
Hiiiii
I am new to this technique. I want to perform a qPCR on my chIP samples.What will be my control for comparing my result and how to calculate.
help me...........
-sssss-
sssss on Jul 3 2010, 01:14 AM said:
Hiiiii
I am new to this technique. I want to perform a qPCR on my chIP samples.What will be my control for comparing my result and how to calculate.
help me...........
I am new to this technique. I want to perform a qPCR on my chIP samples.What will be my control for comparing my result and how to calculate.
help me...........
Hi...first of all you're probably better off posting ChIP questions in the ChIP forum...
Second...I like to have a positive and a negative control for my ChIP...for negative control I use something that shouldn't be bound much by the factor I'm looking at (or has a well known histone modification profile) such as LINE1...for a positive control, you need to search the literature and find a gene that is regulated by the factor your interested in. If your doing ChIP for histones this should be pretty easy to find a good positive control.
As for how to calculate, I typically do %input, there's some old posts in the ChIP section on how to do this I think. Others use fold over IgG, but I find that to be noisy and unreliable, at least for what I do...
MM
-Mighty Mouse-
Mighty Mouse on Jul 5 2010, 06:52 PM said:
sssss on Jul 3 2010, 01:14 AM said:
Hiiiii
I am new to this technique. I want to perform a qPCR on my chIP samples.What will be my control for comparing my result and how to calculate.
help me...........
I am new to this technique. I want to perform a qPCR on my chIP samples.What will be my control for comparing my result and how to calculate.
help me...........
Hi...first of all you're probably better off posting ChIP questions in the ChIP forum...
Second...I like to have a positive and a negative control for my ChIP...for negative control I use something that shouldn't be bound much by the factor I'm looking at (or has a well known histone modification profile) such as LINE1...for a positive control, you need to search the literature and find a gene that is regulated by the factor your interested in. If your doing ChIP for histones this should be pretty easy to find a good positive control.
As for how to calculate, I typically do %input, there's some old posts in the ChIP section on how to do this I think. Others use fold over IgG, but I find that to be noisy and unreliable, at least for what I do...
MM
Thanks for the reply
Can't we take only IgG or Input sample as control
-sssss-
Thanks for the reply
Can't we take only IgG or Input sample as control