Endogenous control validation - (Nov/15/2006 )
Hi all. I am again screening primers for housekeeping genes (in N.benth. leaf tissue).
The last time I did this (for tomato) I tested the candidate housekeeping genes in wild type, empty vector, and in treated plants. I ran the test on three separate occasions with three biological replicates.
The point of all this was establish that the housekeeping genes were not affected by the vector or the treatment.
Are other folks validating their housekeeping genes to this extent? Wondering if this could be accomplished in just one test run.
Yes, it is really very important to check the housekeeping genes are not affected by your treatment.