Hello,

I am concerned with difference among NFs for our experimental setup of
two groups of tissue pools. The NFs are near 0.3 in one group, and 2.5
in the other. In this setup we used 7 references genes, and the
overall M is 0.650.

Using geNorm we selected the two most stable genes among the 7,
obtaining a M of 0.169 with the two. However, the NF histogram looks
about the same as with the 7 ref genes: near 0.3 for one tissue group
and near 2.5 for the other group. What's the difference between M and
NF histogram when assessing ref genes stability?

How can we interpret the final CNRQ histogram for our genes of
interest based on those results about the reference genes? Is this
strong difference among NFs disrupt the normalization, or on the other
hand removes the expression bias detected among the reference genes?
Can we make fold difference assumption in our genes of interest
knowing this difference between NFs?

thanks in advance for any insight you can give me!

Phil